Homotypic Dimerization of a Maltose Kinase for Molecular Scaffolding

Supplementary Information For:

Homotypic dimerization of a maltose kinase for molecular scaffolding

Jun Li1, †, Xiaotao Guan1, †, Neil Shaw1,3, Weiming Chen3, Yu Dong1, Xiaoling Xu4, Xuemei Li1, Zihe Rao1,2,3,*.

1National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Beijing, 100101, China.

2Structure Biology Laboratory, Tsinghua University, Beijing, 100084, China.

3Tianjin Key Laboratory of Protein Science, College of Life Sciences, Nankai University, Tianjin, 300071, China.

4Institute of Aging Research, School of Medicine,Hangzhou Normal University, 310036, China.

*To whom correspondence may be addressed. E-mail:.

†These authors contributed equally to this work.

This document includes –

Supplementary Text

Supplementary Figures S1-S6

Supplementary Tables S1-S3

Supplementary Text

Nucleotide binding site of MtbMaK

Our attempts to crystallize MtbMaK in complex with ATP analogues failed. No electron density for ATP was observed. To gain insights into the ATP-binding site of MtbMaK, we first searched for structures of kinases that could be superimposed over the structure of MtbMaK. Dali search for structural homologues of MtbMaK retrieved prokaryotic as well as eukaryotic kinases as low structural matches. Although alignment of the primary sequences of these structures with MtbMaK revealed no significant conservation over the entire length, some conservation of the primary sequence around the ATP-binding site was observed. Using this clue, structural elements corresponding to the nucleotide-binding site of MtbMaK (111 amino acids) were used as a template to search for structural homologues again. This retrieved the structure of 3', 5" - aminoglycoside phosphotransferase type IIIa (APH; PDB code 1J7U) in complex with ATP analogue AMPPNP as one of the top hits (Fig. 4a). The nucleotide binding site of APH was superimposed over the structure of MtbMaK. This region of APH superimposed over an equivalent region of MtbMaK with an r.m.s.d.of 2.6 Å between the Cα atoms of 90 matching residues. Using such an approach, AMPPNP-Mg of APH could be fitted into the nucleotide-binding site of MtbMaK (Fig. 4b) and a putative mode of ATP binding by MtbMaK could be inferred. The mode of ATP binding was verified by performing alanine-scanning mutagenesis of residues of MtbMaK interacting with the superimposed AMPPNP-Mg moiety (Fig. 4c and Supplementary Table S4).

According to this mode of ATP binding, the adenosine ring of AMPPNP may be held in position by hydrophobic interactions with I155, I338, L329 and Y214. I155A and I338A mutants exhibited only 27% and 36% activity when compared to the wild type enzyme, while the L329A and Y214A mutants retained less than 10% activity of the wild type MaK (Fig. 4c and Supplementary Table S4). The possibility of the adenine moiety binding further deeper into the cavity was refuted by mutating T212 and N173 to alanine. Both these mutations had very little effect on the enzymatic activity of MaK. The ribosyl moiety of AMPPNP possibly interacts with E220. E220A mutant retained nearly 40% of its original activity. In PKA, the corresponding residue, E127, interacts with the ribosyl group of ATP and is important for inducing closure of the lobe. Surprisingly, no glutamate is found in vicinity of the ribose in MTR kinase. The conserved K157 is the only residue that is known to interact directly with both α and β phosphate groups of ATP. Similar to other kinases, the position of its side chain is fixed by a glutamate residue (E341) in MtbMaK. Mutating either K157 or E341to alanine resulted in an almost complete loss in enzymatic activity. In protein kinases, in addition to the lysine, the phosphate groups of ATP are tethered by Mg2+ ions via interactions with two conserved motifs - HXDLKXXN and DFG. Similar motifs - 320HGDLHXXQ327 and 339DFE341–of MtbMaK are in proximity of the superimposed ligands (Fig. 4a) and are likely to participate in binding Mg2+ ions. Specifically,residues D339 and Q327bind Mg2+ (1) and Mg2+ (2). This role of D339 is similar to that of D250 of MTR kinase and D301 of choline kinase located at structurally equivalent positions within a ‘DXE’ motif. D322 is considered as a catalytic residue based on the mechanism of phosphorylation by structurally similar kinases. For instance, in the MTR kinase, D233, which is part of the HGD motif, performs this catalytic role. The aspartate residue polarizes the O1’ atom of maltose for mounting a nucleophilic attack on the γ-phosphate group of ATP. In the binary complex of MtbMaK with maltose, the OD2 atom of D322 is forming a 3.1 Å hydrogen bond with the O1’ atom of maltose. Thus, the 320HGDLHXXQ327 motif is essential for not only binding Mg2+ ions, but is also important for binding maltose and catalysis. The significance of the two conserved 320HGDLHXXQ327 and 339DFE341motifs is underscored by the abolishment of enzymatic activity of D322A, Q327A, D339A and E341A mutants (Fig. 4c and Supplementary Table S4). Strands β8’ and β9’ of MtbMaK are different from other kinases because they originate from the second subunit of the MtbMaK present in the dimer. Usually a Gly-rich motif is found between the two strands in structures of kinases. Surprisingly, instead of a Gly-rich loop, in MtbMaK, an142EQS144loop linking β8’ with β9’ isobserved. Notably, the 142EQS144loop is conserved across all maltose kinase homologues (Supplementary Fig. S3). Amino acids from this loop are likely to play a role in positioning of the phosphate groups of ATP for catalysis via direct interactions or through interactions mediated by Mg2+ ions (Fig. 4b). The conformation of the residues from this loop is different in Chain A of the dimer where a molecule of Bis-Tris is bound in place of maltose. This observation suggests that the 142EQS144loop undergoes conformational changes during binding of the ligands. Mutating either of the residues from this loop to alanine decreased the enzymatic activity dramatically.Interestingly,E142A and S144A mutationsexhibitedgreater loss in activity when compared to the Q143Amutation (Fig. 4c and Supplementary Table S4).A possible explanation for this is that the conformation of the 142EQS144loop is perturbed in E142A and S144A mutants because E142 and S144 are interacting with R21 and R161, respectively. As a result, the side chain of Q143 occupies the Mg2+(1) binding site in these mutants, which is more deleterious to the activity than the removal of the side chain of Q143. Thus, the plausible mode of ATP binding for MtbMaK deduced by superimposing AMPPNP-Mg and performing mutagenesis of residues interacting with the superimposed ATP-analogue suggests a role for 142EQS144loopin binding ATP. Although intriguing, this is not entirely surprising because the 142EQS144 loop occupies a similar position in MtbMaK as the glycine-rich loop of most kinases that is known to tether ATP.

Supplementary Figures S1-S6

Supplementary Figure S1. Schematic diagram of the TreS-MaK-GlgE-GlgB pathway for synthesis of α-glucan.

Cartoon representation of the structures of a tetramer of TreS (PDB code 4LXF), a dimer of MaK (this study; PDB code 4O7O) and a dimer of GlgE (PDB code 1H54) are shown.

Supplementary Figure S2.Dimer of checkpoint kinase 2 (Chk2, PDB code 2CN5).

A cartoon representation of a dimer of Chk2. Dimerization is mediated by the C-lobe and is essential for activation of the kinase. Activation segments (T-loops) are mutually exchanged between monomers of a dimer. ADP bound to the protein is shown as sticks.

Supplementary Figure S3. Sequence alignment of MtbMaK homologues.

Secondary structural elements of MtbMaK are labeled. Absolutely conserved residues are colored with a red background. Residues marked with stars denote conserved residues of the EQSNXS motif, while residues marked with triangles denotes conserved residues in maltose-binding site.

Supplementary Figure S4.Isomers of maltose

Maltose exists in equilibrium between its α and β isomers in solution.

Supplementary Figure S5.HPLC analysis of MtbMaK and enzymaticactivity of protein from different fractions.

(a) Pure MtbMaK consistently eluted in two peaks during HPLC runs using a Nanofilm SEC-250 column (Sepax Technologies, Inc.). Comparison of the elution profile with those of molecular weight standards suggested that recombinant MtbMaK exists as a mixture of dimers and monomers in solution. (b) Kinase activity of MtbMaK eluted in the peaks denoted as dimer and monomer was monitored continuously. The decrease in absorption of NADH at 340 nm denotes the production of ADP (described in Methods). Both, dimeric and monomericforms of MtbMaK were active. Monomeric MtbMaKdisplayed a relatively higher reaction velocity. A control reaction that did not contain MtbMaK was run under identical conditions as the test.

Supplementary Figure S6. Tetramer of MtbMaK

Cartoon representation of a tetramer of MtbMaK. Analysis of the symmetry mates of MtbMaK revealed that MaK could possibly form tetramers. Subsequent PISA analysis of the structure of MtbMaK suggested a high probably (CSS score of 0.769) of MaK forming a biological assembly made up of four units of MaK. Thus, MtbMaK has the ability to oligomerize in solution.

Supplementary Figure S7. Kinetic curve of MtbMaK for maltose

MtbMaK activity was assayed using different concentrations of maltose. This activity was plotted as a function of the concentration of maltose. Error bars represent s.d. (n=3).

Supplementary Tables S1-S3

Supplementary Table S1 List of intermolecular interactions (≤4 Å) between the subunits of MtbMaK. Hydrogen bonds, as suggested by NCONTACT program of CCP4 (6), are highlighted in green, while salt bridges are highlighted in blue.

No. / Chain A / Dist. [Å] / Chain B
1 / A: / ARG / 21 / [ / O / ] / 3.67 / B: / GLN / 143 / [ / CB / ]
2 / A: / ARG / 21 / [ / O / ] / 3.80 / B: / GLN / 143 / [ / OE1 / ]
3 / A: / ARG / 21 / [ / CD / ] / 3.94 / B: / ASP / 140 / [ / CB / ]
4 / A: / ARG / 21 / [ / NE / ] / 3.07 / B: / ASP / 140 / [ / O / ]
5 / A: / ARG / 21 / [ / NE / ] / 3.93 / B: / ASP / 140 / [ / C / ]
6 / A: / ARG / 21 / [ / NE / ] / 3.74 / B: / ASP / 140 / [ / CB / ]
7 / A: / ARG / 21 / [ / CZ / ] / 3.80 / B: / ASP / 140 / [ / O / ]
8 / A: / ARG / 21 / [ / NH2 / ] / 3.65 / B: / ASP / 140 / [ / O / ]
9 / A: / ARG / 21 / [ / NH2 / ] / 3.87 / B: / ALA / 141 / [ / O / ]
10 / A: / TRP / 22 / [ / O / ] / 3.65 / B: / GLN / 143 / [ / O / ]
11 / A: / TRP / 22 / [ / CA / ] / 3.44 / B: / GLN / 143 / [ / O / ]
12 / A: / TRP / 22 / [ / CB / ] / 3.07 / B: / GLN / 143 / [ / O / ]
13 / A: / TRP / 22 / [ / CB / ] / 3.92 / B: / GLN / 143 / [ / C / ]
14 / A: / TRP / 22 / [ / CD1 / ] / 3.88 / B: / THR / 146 / [ / OG1 / ]
15 / A: / TYR / 23 / [ / O / ] / 3.56 / B: / GLN / 143 / [ / OE1 / ]
16 / A: / ALA / 24 / [ / CB / ] / 3.83 / B: / GLN / 143 / [ / NE2 / ]
17 / A: / ARG / 42 / [ / CZ / ] / 3.82 / B: / GLU / 129 / [ / CG / ]
18 / A: / ARG / 42 / [ / NH2 / ] / 3.88 / B: / GLU / 129 / [ / CG / ]
19 / A: / LEU / 94 / [ / O / ] / 3.80 / B: / VAL / 148 / [ / CG2 / ]
20 / A: / LEU / 94 / [ / O / ] / 3.79 / B: / VAL / 138 / [ / CG2 / ]
21 / A: / LEU / 94 / [ / CD1 / ] / 3.83 / B: / THR / 146 / [ / CB / ]
22 / A: / LEU / 94 / [ / CD1 / ] / 3.75 / B: / THR / 146 / [ / CG2 / ]
23 / A: / TYR / 95 / [ / O / ] / 3.98 / B: / ALA / 133 / [ / O / ]
24 / A: / TYR / 95 / [ / O / ] / 3.88 / B: / PHE / 132 / [ / CZ / ]
25 / A: / TYR / 95 / [ / O / ] / 3.77 / B: / ALA / 134 / [ / CA / ]
26 / A: / TYR / 95 / [ / CD1 / ] / 3.97 / B: / VAL / 138 / [ / CG1 / ]
27 / A: / TYR / 95 / [ / CE1 / ] / 3.88 / B: / ASP / 140 / [ / OD2 / ]
28 / A: / TYR / 95 / [ / CE1 / ] / 3.54 / B: / VAL / 138 / [ / CG1 / ]
29 / A: / TYR / 95 / [ / CZ / ] / 3.87 / B: / ASP / 140 / [ / OD1 / ]
30 / A: / TYR / 95 / [ / CZ / ] / 3.87 / B: / VAL / 138 / [ / CG1 / ]
31 / A: / TYR / 95 / [ / OH / ] / 3.49 / B: / ASP / 140 / [ / CG / ]
32 / A: / TYR / 95 / [ / OH / ] / 2.71 / B: / ASP / 140 / [ / OD1 / ]
33 / A: / TYR / 95 / [ / OH / ] / 3.52 / B: / ASP / 140 / [ / OD2 / ]
34 / A: / ASP / 96 / [ / CA / ] / 3.70 / B: / ALA / 133 / [ / O / ]
35 / A: / ASP / 96 / [ / C / ] / 3.80 / B: / ALA / 133 / [ / O / ]
36 / A: / ASP / 96 / [ / CB / ] / 3.78 / B: / PHE / 132 / [ / CD1 / ]
37 / A: / ASP / 96 / [ / CB / ] / 3.62 / B: / PHE / 132 / [ / CE1 / ]
38 / A: / ASP / 96 / [ / OD1 / ] / 3.76 / B: / PHE / 132 / [ / CA / ]
39 / A: / ASP / 96 / [ / OD1 / ] / 3.51 / B: / ALA / 133 / [ / O / ]
40 / A: / ASP / 96 / [ / OD1 / ] / 3.57 / B: / ALA / 133 / [ / N / ]
41 / A: / ASP / 96 / [ / OD1 / ] / 3.99 / B: / PRO / 131 / [ / O / ]
42 / A: / VAL / 97 / [ / N / ] / 3.00 / B: / ALA / 133 / [ / O / ]
43 / A: / VAL / 97 / [ / CA / ] / 3.88 / B: / PRO / 136 / [ / CG / ]
44 / A: / VAL / 97 / [ / CA / ] / 3.90 / B: / PRO / 136 / [ / CD / ]
45 / A: / VAL / 97 / [ / CA / ] / 3.92 / B: / ALA / 133 / [ / O / ]
46 / A: / VAL / 97 / [ / CB / ] / 3.74 / B: / PRO / 136 / [ / CG / ]
47 / A: / VAL / 97 / [ / CB / ] / 3.44 / B: / PRO / 136 / [ / CD / ]
48 / A: / VAL / 97 / [ / CB / ] / 3.68 / B: / ALA / 133 / [ / O / ]
49 / A: / VAL / 97 / [ / CG1 / ] / 3.51 / B: / PRO / 136 / [ / CG / ]
50 / A: / VAL / 97 / [ / CG1 / ] / 3.62 / B: / PRO / 136 / [ / CD / ]
51 / A: / VAL / 97 / [ / CG2 / ] / 3.65 / B: / ALA / 133 / [ / O / ]
52 / A: / PRO / 100 / [ / C / ] / 3.75 / B: / PHE / 150 / [ / CZ / ]
53 / A: / PRO / 100 / [ / O / ] / 3.86 / B: / PHE / 150 / [ / CZ / ]
54 / A: / PRO / 100 / [ / CB / ] / 3.93 / B: / VAL / 148 / [ / CG1 / ]
55 / A: / PRO / 100 / [ / CB / ] / 3.85 / B: / PHE / 150 / [ / CZ / ]
56 / A: / GLN / 101 / [ / N / ] / 3.73 / B: / PHE / 150 / [ / CZ / ]
57 / A: / GLN / 101 / [ / CA / ] / 3.96 / B: / PHE / 150 / [ / CZ / ]
58 / A: / GLN / 101 / [ / C / ] / 3.98 / B: / ASP / 127 / [ / OD1 / ]
59 / A: / GLN / 101 / [ / CG / ] / 3.99 / B: / ASP / 127 / [ / CG / ]
60 / A: / GLN / 101 / [ / CG / ] / 3.36 / B: / ASP / 127 / [ / OD1 / ]
61 / A: / GLN / 101 / [ / CG / ] / 3.78 / B: / ASP / 127 / [ / OD2 / ]
62 / A: / GLN / 101 / [ / CD / ] / 3.72 / B: / ARG / 186 / [ / NH2 / ]
63 / A: / GLN / 101 / [ / OE1 / ] / 3.82 / B: / ARG / 152 / [ / NH1 / ]
64 / A: / GLN / 101 / [ / OE1 / ] / 3.90 / B: / ARG / 186 / [ / CZ / ]
65 / A: / GLN / 101 / [ / OE1 / ] / 3.83 / B: / ARG / 186 / [ / NH2 / ]
66 / A: / GLN / 101 / [ / OE1 / ] / 3.93 / B: / ARG / 186 / [ / NE / ]
67 / A: / GLN / 101 / [ / NE2 / ] / 3.68 / B: / ARG / 152 / [ / NH1 / ]
68 / A: / GLN / 101 / [ / NE2 / ] / 3.34 / B: / ARG / 186 / [ / NH2 / ]
69 / A: / PHE / 102 / [ / N / ] / 3.80 / B: / ASP / 127 / [ / OD1 / ]
70 / A: / PHE / 102 / [ / CA / ] / 3.75 / B: / ASP / 127 / [ / OD1 / ]
71 / A: / LEU / 104 / [ / CD2 / ] / 3.95 / B: / PHE / 150 / [ / CD2 / ]
72 / A: / SER / 105 / [ / OG / ] / 3.95 / B: / ASP / 127 / [ / CG / ]
73 / A: / VAL / 108 / [ / O / ] / 3.82 / B: / VAL / 108 / [ / CG1 / ]
74 / A: / SER / 109 / [ / CA / ] / 3.99 / B: / VAL / 108 / [ / O / ]
75 / A: / SER / 109 / [ / CB / ] / 3.90 / B: / VAL / 108 / [ / O / ]
76 / A: / SER / 109 / [ / CB / ] / 3.49 / B: / ARG / 124 / [ / NE / ]
77 / A: / SER / 109 / [ / CB / ] / 3.87 / B: / ARG / 124 / [ / CZ / ]
78 / A: / SER / 109 / [ / OG / ] / 3.93 / B: / ARG / 124 / [ / NE / ]
79 / A: / SER / 109 / [ / OG / ] / 3.88 / B: / ARG / 124 / [ / CZ / ]
80 / A: / SER / 109 / [ / OG / ] / 3.67 / B: / ARG / 124 / [ / NH2 / ]
81 / A: / ARG / 124 / [ / CZ / ] / 3.54 / B: / SER / 109 / [ / CB / ]
82 / A: / ARG / 124 / [ / NH1 / ] / 3.75 / B: / SER / 109 / [ / CB / ]
83 / A: / ARG / 124 / [ / NH2 / ] / 3.48 / B: / SER / 109 / [ / CB / ]
84 / A: / ASP / 127 / [ / CB / ] / 3.77 / B: / SER / 105 / [ / OG / ]
85 / A: / ASP / 127 / [ / CG / ] / 3.74 / B: / SER / 105 / [ / OG / ]
86 / A: / ASP / 127 / [ / OD1 / ] / 3.12 / B: / ARG / 42 / [ / NH1 / ]
87 / A: / ASP / 127 / [ / OD1 / ] / 3.65 / B: / PHE / 102 / [ / CD2 / ]
88 / A: / ASP / 127 / [ / OD2 / ] / 3.95 / B: / GLN / 101 / [ / CG / ]
89 / A: / ASP / 127 / [ / OD2 / ] / 3.40 / B: / SER / 105 / [ / OG / ]
90 / A: / ASP / 127 / [ / OD2 / ] / 3.47 / B: / GLN / 101 / [ / NE2 / ]
91 / A: / GLU / 129 / [ / CG / ] / 3.88 / B: / ARG / 42 / [ / NE / ]
92 / A: / GLU / 129 / [ / CG / ] / 3.55 / B: / ARG / 42 / [ / CZ / ]
93 / A: / GLU / 129 / [ / CG / ] / 3.96 / B: / ARG / 42 / [ / NH1 / ]
94 / A: / GLU / 129 / [ / CG / ] / 3.54 / B: / ARG / 42 / [ / NH2 / ]
95 / A: / GLU / 129 / [ / CD / ] / 3.89 / B: / ARG / 42 / [ / CZ / ]
96 / A: / GLU / 129 / [ / CD / ] / 3.28 / B: / ASP / 69 / [ / OD2 / ]
97 / A: / GLU / 129 / [ / CD / ] / 3.36 / B: / ARG / 42 / [ / NH2 / ]
98 / A: / GLU / 129 / [ / OE1 / ] / 3.84 / B: / ASP / 69 / [ / OD1 / ]
99 / A: / GLU / 129 / [ / OE1 / ] / 3.21 / B: / ASP / 69 / [ / CG / ]
100 / A: / GLU / 129 / [ / OE1 / ] / 2.08 / B: / ASP / 69 / [ / OD2 / ]
101 / A: / GLU / 129 / [ / OE1 / ] / 3.89 / B: / ARG / 42 / [ / NH2 / ]
102 / A: / GLU / 129 / [ / OE2 / ] / 3.38 / B: / ARG / 42 / [ / NH2 / ]
103 / A: / GLU / 129 / [ / OE2 / ] / 3.51 / B: / ALA / 98 / [ / CB / ]
104 / A: / LEU / 130 / [ / O / ] / 3.87 / B: / ALA / 98 / [ / CB / ]
105 / A: / PHE / 132 / [ / CA / ] / 3.57 / B: / ASP / 96 / [ / OD1 / ]
106 / A: / PHE / 132 / [ / C / ] / 3.94 / B: / ASP / 96 / [ / OD1 / ]
107 / A: / PHE / 132 / [ / CB / ] / 3.79 / B: / PRO / 72 / [ / CD / ]
108 / A: / PHE / 132 / [ / CG / ] / 3.71 / B: / PRO / 72 / [ / CD / ]
109 / A: / PHE / 132 / [ / CD1 / ] / 3.90 / B: / GLU / 71 / [ / CG / ]
110 / A: / PHE / 132 / [ / CD1 / ] / 3.43 / B: / GLU / 71 / [ / CD / ]
111 / A: / PHE / 132 / [ / CD1 / ] / 3.63 / B: / GLU / 71 / [ / OE1 / ]
112 / A: / PHE / 132 / [ / CD1 / ] / 3.55 / B: / GLU / 71 / [ / OE2 / ]
113 / A: / PHE / 132 / [ / CD1 / ] / 3.78 / B: / ASP / 96 / [ / CB / ]
114 / A: / PHE / 132 / [ / CD1 / ] / 3.63 / B: / ASP / 96 / [ / CG / ]
115 / A: / PHE / 132 / [ / CD1 / ] / 3.60 / B: / ASP / 96 / [ / OD1 / ]
116 / A: / PHE / 132 / [ / CD2 / ] / 3.69 / B: / PRO / 72 / [ / CG / ]
117 / A: / PHE / 132 / [ / CD2 / ] / 3.41 / B: / PRO / 72 / [ / CD / ]
118 / A: / PHE / 132 / [ / CE1 / ] / 3.73 / B: / GLU / 71 / [ / CD / ]
119 / A: / PHE / 132 / [ / CE1 / ] / 3.91 / B: / GLU / 71 / [ / OE1 / ]
120 / A: / PHE / 132 / [ / CE1 / ] / 3.44 / B: / GLU / 71 / [ / OE2 / ]
121 / A: / PHE / 132 / [ / CE1 / ] / 3.90 / B: / ASP / 96 / [ / CB / ]
122 / A: / PHE / 132 / [ / CE1 / ] / 3.59 / B: / TYR / 95 / [ / O / ]
123 / A: / PHE / 132 / [ / CE1 / ] / 3.86 / B: / ASP / 96 / [ / CA / ]
124 / A: / PHE / 132 / [ / CE2 / ] / 3.84 / B: / GLU / 75 / [ / CG / ]
125 / A: / PHE / 132 / [ / CE2 / ] / 3.87 / B: / GLU / 75 / [ / CD / ]
126 / A: / PHE / 132 / [ / CZ / ] / 3.84 / B: / GLU / 75 / [ / CG / ]
127 / A: / PHE / 132 / [ / CZ / ] / 3.80 / B: / GLU / 75 / [ / CD / ]
128 / A: / PHE / 132 / [ / CZ / ] / 3.71 / B: / GLU / 75 / [ / OE1 / ]
129 / A: / PHE / 132 / [ / CZ / ] / 3.74 / B: / TYR / 95 / [ / O / ]
130 / A: / ALA / 133 / [ / O / ] / 3.67 / B: / TYR / 95 / [ / O / ]
131 / A: / ALA / 133 / [ / O / ] / 3.62 / B: / ASP / 96 / [ / CA / ]
132 / A: / ALA / 133 / [ / O / ] / 3.66 / B: / ASP / 96 / [ / OD1 / ]
133 / A: / ALA / 133 / [ / O / ] / 3.66 / B: / ASP / 96 / [ / C / ]
134 / A: / ALA / 133 / [ / O / ] / 2.81 / B: / VAL / 97 / [ / N / ]
135 / A: / ALA / 133 / [ / O / ] / 3.76 / B: / VAL / 97 / [ / CA / ]
136 / A: / ALA / 133 / [ / O / ] / 3.63 / B: / VAL / 97 / [ / CB / ]
137 / A: / ALA / 133 / [ / O / ] / 3.24 / B: / VAL / 97 / [ / CG2 / ]
138 / A: / ALA / 133 / [ / N / ] / 3.30 / B: / ASP / 96 / [ / OD1 / ]
139 / A: / ALA / 133 / [ / C / ] / 4.00 / B: / VAL / 97 / [ / N / ]
140 / A: / ALA / 133 / [ / C / ] / 3.93 / B: / VAL / 97 / [ / CG2 / ]
141 / A: / ALA / 133 / [ / CB / ] / 3.85 / B: / VAL / 97 / [ / CG2 / ]
142 / A: / ALA / 134 / [ / O / ] / 3.64 / B: / VAL / 97 / [ / CG2 / ]
143 / A: / ALA / 134 / [ / CA / ] / 3.71 / B: / TYR / 95 / [ / O / ]
144 / A: / ALA / 134 / [ / C / ] / 3.82 / B: / TYR / 95 / [ / O / ]
145 / A: / ALA / 134 / [ / C / ] / 3.94 / B: / VAL / 97 / [ / CG2 / ]
146 / A: / GLN / 135 / [ / N / ] / 3.88 / B: / TYR / 95 / [ / C / ]
147 / A: / GLN / 135 / [ / N / ] / 2.99 / B: / TYR / 95 / [ / O / ]
148 / A: / GLN / 135 / [ / CA / ] / 3.80 / B: / TYR / 95 / [ / O / ]
149 / A: / GLN / 135 / [ / CA / ] / 3.65 / B: / VAL / 97 / [ / CG2 / ]
150 / A: / GLN / 135 / [ / CB / ] / 3.58 / B: / TYR / 95 / [ / C / ]
151 / A: / GLN / 135 / [ / CB / ] / 3.48 / B: / TYR / 95 / [ / O / ]
152 / A: / GLN / 135 / [ / CB / ] / 3.51 / B: / LEU / 94 / [ / O / ]
153 / A: / GLN / 135 / [ / CG / ] / 3.47 / B: / LEU / 94 / [ / O / ]
154 / A: / GLN / 135 / [ / CD / ] / 3.77 / B: / PRO / 100 / [ / CB / ]
155 / A: / GLN / 135 / [ / CD / ] / 3.20 / B: / PRO / 100 / [ / CG / ]
156 / A: / GLN / 135 / [ / CD / ] / 3.94 / B: / LEU / 94 / [ / O / ]
157 / A: / GLN / 135 / [ / OE1 / ] / 3.58 / B: / PRO / 100 / [ / CB / ]
158 / A: / GLN / 135 / [ / OE1 / ] / 3.49 / B: / PRO / 100 / [ / CG / ]
159 / A: / GLN / 135 / [ / NE2 / ] / 3.15 / B: / ASP / 96 / [ / O / ]
160 / A: / GLN / 135 / [ / NE2 / ] / 3.83 / B: / VAL / 97 / [ / O / ]
161 / A: / GLN / 135 / [ / NE2 / ] / 3.44 / B: / PRO / 100 / [ / CB / ]
162 / A: / GLN / 135 / [ / NE2 / ] / 2.53 / B: / PRO / 100 / [ / CG / ]
163 / A: / GLN / 135 / [ / NE2 / ] / 3.71 / B: / PRO / 100 / [ / CD / ]
164 / A: / GLN / 135 / [ / NE2 / ] / 3.46 / B: / ASP / 96 / [ / C / ]
165 / A: / GLN / 135 / [ / NE2 / ] / 3.45 / B: / VAL / 97 / [ / N / ]
166 / A: / GLN / 135 / [ / NE2 / ] / 3.11 / B: / VAL / 97 / [ / CA / ]
167 / A: / GLN / 135 / [ / NE2 / ] / 3.88 / B: / VAL / 97 / [ / C / ]
168 / A: / PRO / 136 / [ / CG / ] / 3.79 / B: / VAL / 97 / [ / CG2 / ]
169 / A: / PRO / 136 / [ / CD / ] / 3.33 / B: / VAL / 97 / [ / CG2 / ]
170 / A: / ARG / 137 / [ / NH1 / ] / 3.89 / B: / ARG / 153 / [ / NH2 / ]
171 / A: / VAL / 138 / [ / CG1 / ] / 3.68 / B: / TYR / 95 / [ / CE1 / ]
172 / A: / VAL / 138 / [ / CG2 / ] / 3.37 / B: / LEU / 94 / [ / O / ]
173 / A: / ASP / 140 / [ / CB / ] / 3.79 / B: / ARG / 21 / [ / CD / ]
174 / A: / ASP / 140 / [ / CG / ] / 3.91 / B: / TRP / 22 / [ / CD1 / ]
175 / A: / ASP / 140 / [ / CG / ] / 3.93 / B: / ARG / 21 / [ / CD / ]
176 / A: / ASP / 140 / [ / OD2 / ] / 2.91 / B: / TRP / 22 / [ / CD1 / ]
177 / A: / ASP / 140 / [ / OD2 / ] / 3.95 / B: / TRP / 22 / [ / NE1 / ]
178 / A: / ASP / 140 / [ / OD2 / ] / 3.92 / B: / TRP / 22 / [ / CB / ]
179 / A: / ASP / 140 / [ / OD2 / ] / 3.77 / B: / TRP / 22 / [ / CG / ]
180 / A: / GLU / 142 / [ / OE1 / ] / 3.04 / B: / ARG / 21 / [ / O / ]
181 / A: / GLU / 142 / [ / OE2 / ] / 3.47 / B: / ALA / 24 / [ / CB / ]
182 / A: / GLN / 143 / [ / CB / ] / 3.98 / B: / PRO / 344 / [ / CB / ]
183 / A: / GLN / 143 / [ / CD / ] / 3.81 / B: / PRO / 344 / [ / CG / ]
184 / A: / GLN / 143 / [ / OE1 / ] / 3.47 / B: / ASP / 339 / [ / OD2 / ]
185 / A: / GLN / 143 / [ / NE2 / ] / 3.84 / B: / GLU / 341 / [ / CG / ]
186 / A: / GLN / 143 / [ / NE2 / ] / 3.90 / B: / ASP / 339 / [ / OD1 / ]
187 / A: / GLN / 143 / [ / NE2 / ] / 3.57 / B: / GLU / 341 / [ / CB / ]
188 / A: / GLN / 143 / [ / NE2 / ] / 3.87 / B: / PRO / 344 / [ / CG / ]
189 / A: / GLN / 143 / [ / NE2 / ] / 3.14 / B: / GLY / 342 / [ / O / ]
190 / A: / SER / 144 / [ / O / ] / 3.17 / B: / ARG / 160 / [ / N / ]
191 / A: / SER / 144 / [ / O / ] / 3.93 / B: / ARG / 160 / [ / CA / ]
192 / A: / SER / 144 / [ / O / ] / 3.73 / B: / ARG / 160 / [ / CB / ]
193 / A: / SER / 144 / [ / O / ] / 3.95 / B: / ARG / 161 / [ / N / ]
194 / A: / SER / 144 / [ / CB / ] / 3.18 / B: / TRP / 22 / [ / O / ]
195 / A: / SER / 144 / [ / CB / ] / 3.74 / B: / TRP / 22 / [ / CA / ]
196 / A: / SER / 144 / [ / CB / ] / 3.88 / B: / TRP / 22 / [ / C / ]
197 / A: / SER / 144 / [ / CB / ] / 3.43 / B: / TRP / 22 / [ / CB / ]
198 / A: / SER / 144 / [ / OG / ] / 3.65 / B: / ARG / 160 / [ / CB / ]
199 / A: / SER / 144 / [ / OG / ] / 3.38 / B: / TRP / 22 / [ / O / ]
200 / A: / SER / 144 / [ / OG / ] / 3.06 / B: / ARG / 161 / [ / N / ]
201 / A: / SER / 144 / [ / OG / ] / 3.75 / B: / ARG / 161 / [ / CA / ]
202 / A: / SER / 144 / [ / OG / ] / 3.31 / B: / ARG / 161 / [ / CB / ]
203 / A: / SER / 144 / [ / OG / ] / 3.90 / B: / ARG / 161 / [ / NH2 / ]
204 / A: / ASN / 145 / [ / CA / ] / 3.24 / B: / VAL / 158 / [ / O / ]
205 / A: / ASN / 145 / [ / CA / ] / 3.92 / B: / ARG / 160 / [ / CG / ]
206 / A: / ASN / 145 / [ / C / ] / 3.54 / B: / VAL / 158 / [ / O / ]
207 / A: / ASN / 145 / [ / CB / ] / 3.33 / B: / VAL / 158 / [ / O / ]
208 / A: / ASN / 145 / [ / CG / ] / 3.64 / B: / LYS / 157 / [ / CD / ]
209 / A: / ASN / 145 / [ / OD1 / ] / 3.57 / B: / LYS / 157 / [ / CD / ]
210 / A: / ASN / 145 / [ / ND2 / ] / 3.99 / B: / LYS / 157 / [ / CD / ]
211 / A: / THR / 146 / [ / O / ] / 3.74 / B: / LEU / 156 / [ / O / ]
212 / A: / THR / 146 / [ / O / ] / 3.53 / B: / VAL / 158 / [ / O / ]
213 / A: / THR / 146 / [ / O / ] / 3.57 / B: / VAL / 158 / [ / CB / ]
214 / A: / THR / 146 / [ / O / ] / 3.72 / B: / LYS / 157 / [ / CA / ]
215 / A: / THR / 146 / [ / O / ] / 3.73 / B: / LYS / 157 / [ / C / ]
216 / A: / THR / 146 / [ / O / ] / 2.80 / B: / VAL / 158 / [ / N / ]
217 / A: / THR / 146 / [ / O / ] / 3.60 / B: / VAL / 158 / [ / CA / ]
218 / A: / THR / 146 / [ / O / ] / 3.99 / B: / VAL / 158 / [ / C / ]
219 / A: / THR / 146 / [ / N / ] / 2.88 / B: / VAL / 158 / [ / O / ]
220 / A: / THR / 146 / [ / CA / ] / 3.96 / B: / VAL / 158 / [ / O / ]
221 / A: / THR / 146 / [ / C / ] / 3.94 / B: / VAL / 158 / [ / N / ]
222 / A: / THR / 146 / [ / CB / ] / 3.97 / B: / LEU / 94 / [ / CD1 / ]
223 / A: / THR / 146 / [ / CB / ] / 3.92 / B: / ARG / 160 / [ / NH2 / ]
224 / A: / THR / 146 / [ / CB / ] / 3.75 / B: / ARG / 160 / [ / NE / ]
225 / A: / THR / 146 / [ / OG1 / ] / 3.89 / B: / ARG / 160 / [ / CD / ]
226 / A: / THR / 146 / [ / OG1 / ] / 3.72 / B: / TRP / 22 / [ / CD1 / ]
227 / A: / THR / 146 / [ / OG1 / ] / 3.92 / B: / TRP / 22 / [ / NE1 / ]
228 / A: / THR / 146 / [ / OG1 / ] / 3.50 / B: / ARG / 160 / [ / CZ / ]
229 / A: / THR / 146 / [ / OG1 / ] / 3.30 / B: / ARG / 160 / [ / NH2 / ]
230 / A: / THR / 146 / [ / OG1 / ] / 3.85 / B: / ARG / 160 / [ / CG / ]
231 / A: / THR / 146 / [ / OG1 / ] / 2.85 / B: / ARG / 160 / [ / NE / ]
232 / A: / THR / 146 / [ / CG2 / ] / 3.97 / B: / LEU / 94 / [ / CD1 / ]
233 / A: / SER / 147 / [ / CA / ] / 3.14 / B: / LEU / 156 / [ / O / ]
234 / A: / SER / 147 / [ / C / ] / 3.44 / B: / LEU / 156 / [ / O / ]
235 / A: / SER / 147 / [ / CB / ] / 3.75 / B: / LEU / 156 / [ / O / ]
236 / A: / SER / 147 / [ / CB / ] / 3.94 / B: / ILE / 155 / [ / CG2 / ]
237 / A: / SER / 147 / [ / OG / ] / 3.76 / B: / LYS / 157 / [ / CA / ]
238 / A: / SER / 147 / [ / OG / ] / 3.64 / B: / LYS / 157 / [ / CB / ]
239 / A: / SER / 147 / [ / OG / ] / 3.97 / B: / LYS / 157 / [ / CG / ]
240 / A: / SER / 147 / [ / OG / ] / 3.78 / B: / LYS / 157 / [ / CD / ]
241 / A: / VAL / 148 / [ / O / ] / 3.70 / B: / LEU / 156 / [ / O / ]
242 / A: / VAL / 148 / [ / O / ] / 3.92 / B: / LEU / 156 / [ / CA / ]
243 / A: / VAL / 148 / [ / O / ] / 3.63 / B: / ALA / 154 / [ / O / ]
244 / A: / VAL / 148 / [ / O / ] / 3.46 / B: / ILE / 155 / [ / CA / ]
245 / A: / VAL / 148 / [ / O / ] / 3.69 / B: / ILE / 155 / [ / C / ]
246 / A: / VAL / 148 / [ / O / ] / 2.93 / B: / LEU / 156 / [ / N / ]
247 / A: / VAL / 148 / [ / N / ] / 2.81 / B: / LEU / 156 / [ / O / ]
248 / A: / VAL / 148 / [ / N / ] / 3.95 / B: / LEU / 156 / [ / C / ]
249 / A: / VAL / 148 / [ / CA / ] / 3.95 / B: / LEU / 156 / [ / O / ]
250 / A: / VAL / 148 / [ / CG1 / ] / 3.82 / B: / PRO / 100 / [ / CB / ]
251 / A: / VAL / 148 / [ / CG1 / ] / 3.92 / B: / LEU / 156 / [ / CB / ]
252 / A: / VAL / 148 / [ / CG2 / ] / 3.88 / B: / LEU / 94 / [ / O / ]
253 / A: / ILE / 149 / [ / CA / ] / 3.51 / B: / ALA / 154 / [ / O / ]
254 / A: / ILE / 149 / [ / C / ] / 3.65 / B: / ALA / 154 / [ / O / ]
255 / A: / ILE / 149 / [ / CG1 / ] / 3.81 / B: / ILE / 155 / [ / CD1 / ]
256 / A: / ILE / 149 / [ / CG2 / ] / 3.33 / B: / ARG / 153 / [ / NE / ]
257 / A: / ILE / 149 / [ / CG2 / ] / 3.57 / B: / ARG / 153 / [ / CZ / ]
258 / A: / ILE / 149 / [ / CG2 / ] / 3.68 / B: / ARG / 153 / [ / NH2 / ]
259 / A: / ILE / 149 / [ / CG2 / ] / 3.99 / B: / ARG / 153 / [ / CD / ]
260 / A: / PHE / 150 / [ / O / ] / 3.48 / B: / ARG / 152 / [ / O / ]
261 / A: / PHE / 150 / [ / O / ] / 3.61 / B: / ALA / 154 / [ / O / ]
262 / A: / PHE / 150 / [ / O / ] / 3.52 / B: / ARG / 153 / [ / C / ]
263 / A: / PHE / 150 / [ / O / ] / 2.75 / B: / ALA / 154 / [ / N / ]
264 / A: / PHE / 150 / [ / O / ] / 3.77 / B: / ALA / 154 / [ / CA / ]
265 / A: / PHE / 150 / [ / O / ] / 3.98 / B: / ALA / 154 / [ / CB / ]
266 / A: / PHE / 150 / [ / O / ] / 3.37 / B: / ARG / 153 / [ / CA / ]
267 / A: / PHE / 150 / [ / N / ] / 2.83 / B: / ALA / 154 / [ / O / ]
268 / A: / PHE / 150 / [ / N / ] / 3.97 / B: / ALA / 154 / [ / C / ]
269 / A: / PHE / 150 / [ / CA / ] / 3.77 / B: / ALA / 154 / [ / O / ]
270 / A: / PHE / 150 / [ / C / ] / 3.89 / B: / ALA / 154 / [ / N / ]
271 / A: / PHE / 150 / [ / CB / ] / 3.94 / B: / ALA / 154 / [ / O / ]
272 / A: / PHE / 150 / [ / CD2 / ] / 3.96 / B: / ALA / 154 / [ / O / ]
273 / A: / PHE / 150 / [ / CE2 / ] / 3.68 / B: / LEU / 104 / [ / CD1 / ]
274 / A: / PHE / 150 / [ / CZ / ] / 3.60 / B: / PRO / 100 / [ / CB / ]
275 / A: / PHE / 150 / [ / CZ / ] / 3.73 / B: / PRO / 100 / [ / C / ]
276 / A: / PHE / 150 / [ / CZ / ] / 3.94 / B: / PRO / 100 / [ / O / ]
277 / A: / PHE / 150 / [ / CZ / ] / 3.76 / B: / GLN / 101 / [ / N / ]
278 / A: / ASP / 151 / [ / CA / ] / 3.57 / B: / ARG / 152 / [ / O / ]
279 / A: / ASP / 151 / [ / C / ] / 3.79 / B: / ARG / 152 / [ / O / ]
280 / A: / ASP / 151 / [ / CG / ] / 3.58 / B: / ARG / 153 / [ / NE / ]
281 / A: / ASP / 151 / [ / CG / ] / 4.00 / B: / ARG / 153 / [ / CZ / ]
282 / A: / ASP / 151 / [ / CG / ] / 3.46 / B: / ARG / 153 / [ / NH2 / ]
283 / A: / ASP / 151 / [ / OD1 / ] / 2.69 / B: / ARG / 153 / [ / NE / ]
284 / A: / ASP / 151 / [ / OD1 / ] / 3.45 / B: / ARG / 153 / [ / CZ / ]
285 / A: / ASP / 151 / [ / OD1 / ] / 3.35 / B: / ARG / 153 / [ / NH2 / ]
286 / A: / ASP / 151 / [ / OD1 / ] / 3.64 / B: / ARG / 153 / [ / CA / ]
287 / A: / ASP / 151 / [ / OD1 / ] / 3.37 / B: / ARG / 153 / [ / CB / ]
288 / A: / ASP / 151 / [ / OD1 / ] / 3.55 / B: / ARG / 153 / [ / CG / ]
289 / A: / ASP / 151 / [ / OD1 / ] / 3.67 / B: / ARG / 153 / [ / CD / ]
290 / A: / ASP / 151 / [ / OD2 / ] / 3.68 / B: / ARG / 153 / [ / NE / ]
291 / A: / ASP / 151 / [ / OD2 / ] / 3.68 / B: / ARG / 153 / [ / CZ / ]
292 / A: / ASP / 151 / [ / OD2 / ] / 2.80 / B: / ARG / 153 / [ / NH2 / ]
293 / A: / ARG / 152 / [ / O / ] / 3.39 / B: / ARG / 152 / [ / O / ]
294 / A: / ARG / 152 / [ / O / ] / 3.13 / B: / ARG / 152 / [ / N / ]
295 / A: / ARG / 152 / [ / O / ] / 3.67 / B: / PHE / 150 / [ / O / ]
296 / A: / ARG / 152 / [ / O / ] / 3.72 / B: / ASP / 151 / [ / CA / ]
297 / A: / ARG / 152 / [ / O / ] / 3.89 / B: / ASP / 151 / [ / C / ]
298 / A: / ARG / 152 / [ / N / ] / 3.08 / B: / ARG / 152 / [ / O / ]
299 / A: / ARG / 152 / [ / NH2 / ] / 3.53 / B: / GLN / 101 / [ / OE1 / ]
300 / A: / ARG / 153 / [ / CA / ] / 3.45 / B: / PHE / 150 / [ / O / ]
301 / A: / ARG / 153 / [ / CA / ] / 3.71 / B: / ASP / 151 / [ / OD1 / ]
302 / A: / ARG / 153 / [ / C / ] / 3.64 / B: / PHE / 150 / [ / O / ]
303 / A: / ARG / 153 / [ / CB / ] / 3.44 / B: / ASP / 151 / [ / OD1 / ]
304 / A: / ARG / 153 / [ / CG / ] / 3.85 / B: / ILE / 149 / [ / CG2 / ]
305 / A: / ARG / 153 / [ / CG / ] / 3.48 / B: / ASP / 151 / [ / OD1 / ]
306 / A: / ARG / 153 / [ / CD / ] / 3.79 / B: / ILE / 149 / [ / CG2 / ]
307 / A: / ARG / 153 / [ / CD / ] / 3.64 / B: / ASP / 151 / [ / OD1 / ]
308 / A: / ARG / 153 / [ / NE / ] / 3.68 / B: / ILE / 149 / [ / CG2 / ]
309 / A: / ARG / 153 / [ / NE / ] / 2.70 / B: / ASP / 151 / [ / OD1 / ]
310 / A: / ARG / 153 / [ / NE / ] / 3.56 / B: / ASP / 151 / [ / CG / ]
311 / A: / ARG / 153 / [ / NE / ] / 3.66 / B: / ASP / 151 / [ / OD2 / ]
312 / A: / ARG / 153 / [ / CZ / ] / 3.81 / B: / ILE / 149 / [ / CG2 / ]
313 / A: / ARG / 153 / [ / CZ / ] / 3.39 / B: / ASP / 151 / [ / OD1 / ]
314 / A: / ARG / 153 / [ / CZ / ] / 3.86 / B: / ASP / 151 / [ / CG / ]
315 / A: / ARG / 153 / [ / CZ / ] / 3.50 / B: / ASP / 151 / [ / OD2 / ]
316 / A: / ARG / 153 / [ / NH1 / ] / 3.99 / B: / ILE / 149 / [ / CG2 / ]
317 / A: / ARG / 153 / [ / NH1 / ] / 3.72 / B: / ARG / 137 / [ / NH2 / ]
318 / A: / ARG / 153 / [ / NH2 / ] / 3.35 / B: / ASP / 151 / [ / OD1 / ]
319 / A: / ARG / 153 / [ / NH2 / ] / 3.41 / B: / ASP / 151 / [ / CG / ]
320 / A: / ARG / 153 / [ / NH2 / ] / 2.72 / B: / ASP / 151 / [ / OD2 / ]
321 / A: / ALA / 154 / [ / O / ] / 3.61 / B: / VAL / 148 / [ / O / ]
322 / A: / ALA / 154 / [ / O / ] / 3.69 / B: / ILE / 149 / [ / CA / ]
323 / A: / ALA / 154 / [ / O / ] / 3.82 / B: / ILE / 149 / [ / C / ]
324 / A: / ALA / 154 / [ / O / ] / 2.94 / B: / PHE / 150 / [ / N / ]
325 / A: / ALA / 154 / [ / O / ] / 3.71 / B: / PHE / 150 / [ / CB / ]
326 / A: / ALA / 154 / [ / O / ] / 3.77 / B: / PHE / 150 / [ / CD2 / ]
327 / A: / ALA / 154 / [ / O / ] / 3.67 / B: / PHE / 150 / [ / O / ]
328 / A: / ALA / 154 / [ / O / ] / 3.77 / B: / PHE / 150 / [ / CA / ]
329 / A: / ALA / 154 / [ / N / ] / 2.88 / B: / PHE / 150 / [ / O / ]
330 / A: / ALA / 154 / [ / CA / ] / 3.91 / B: / PHE / 150 / [ / O / ]
331 / A: / ILE / 155 / [ / CA / ] / 3.51 / B: / VAL / 148 / [ / O / ]
332 / A: / ILE / 155 / [ / C / ] / 3.73 / B: / VAL / 148 / [ / O / ]
333 / A: / ILE / 155 / [ / CD1 / ] / 3.55 / B: / ILE / 149 / [ / CG1 / ]
334 / A: / LEU / 156 / [ / O / ] / 3.79 / B: / SER / 147 / [ / CB / ]
335 / A: / LEU / 156 / [ / O / ] / 3.85 / B: / SER / 147 / [ / OG / ]
336 / A: / LEU / 156 / [ / O / ] / 3.88 / B: / THR / 146 / [ / O / ]
337 / A: / LEU / 156 / [ / O / ] / 3.24 / B: / SER / 147 / [ / CA / ]
338 / A: / LEU / 156 / [ / O / ] / 3.53 / B: / SER / 147 / [ / C / ]
339 / A: / LEU / 156 / [ / O / ] / 2.84 / B: / VAL / 148 / [ / N / ]
340 / A: / LEU / 156 / [ / O / ] / 3.74 / B: / VAL / 148 / [ / O / ]
341 / A: / LEU / 156 / [ / O / ] / 3.91 / B: / VAL / 148 / [ / CA / ]
342 / A: / LEU / 156 / [ / N / ] / 2.95 / B: / VAL / 148 / [ / O / ]
343 / A: / LEU / 156 / [ / CA / ] / 3.89 / B: / VAL / 148 / [ / O / ]
344 / A: / LEU / 156 / [ / C / ] / 3.98 / B: / VAL / 148 / [ / N / ]
345 / A: / LEU / 156 / [ / CB / ] / 3.96 / B: / VAL / 148 / [ / O / ]
346 / A: / LYS / 157 / [ / CA / ] / 3.74 / B: / SER / 147 / [ / OG / ]
347 / A: / LYS / 157 / [ / CA / ] / 3.79 / B: / THR / 146 / [ / O / ]
348 / A: / LYS / 157 / [ / C / ] / 3.83 / B: / THR / 146 / [ / O / ]
349 / A: / LYS / 157 / [ / CB / ] / 3.74 / B: / SER / 147 / [ / OG / ]
350 / A: / LYS / 157 / [ / CG / ] / 4.00 / B: / ASN / 145 / [ / OD1 / ]
351 / A: / LYS / 157 / [ / CG / ] / 3.72 / B: / SER / 147 / [ / OG / ]
352 / A: / LYS / 157 / [ / CE / ] / 3.75 / B: / ASN / 145 / [ / CG / ]
353 / A: / LYS / 157 / [ / CE / ] / 3.57 / B: / ASN / 145 / [ / ND2 / ]
354 / A: / LYS / 157 / [ / NZ / ] / 3.79 / B: / ASN / 145 / [ / ND2 / ]
355 / A: / VAL / 158 / [ / O / ] / 3.30 / B: / ASN / 145 / [ / CA / ]
356 / A: / VAL / 158 / [ / O / ] / 3.25 / B: / ASN / 145 / [ / CB / ]
357 / A: / VAL / 158 / [ / O / ] / 3.06 / B: / THR / 146 / [ / N / ]
358 / A: / VAL / 158 / [ / O / ] / 3.69 / B: / ASN / 145 / [ / C / ]
359 / A: / VAL / 158 / [ / O / ] / 3.52 / B: / THR / 146 / [ / O / ]
360 / A: / VAL / 158 / [ / N / ] / 2.90 / B: / THR / 146 / [ / O / ]
361 / A: / VAL / 158 / [ / CA / ] / 3.71 / B: / THR / 146 / [ / O / ]
362 / A: / VAL / 158 / [ / C / ] / 3.92 / B: / ASN / 145 / [ / CB / ]
363 / A: / VAL / 158 / [ / C / ] / 4.00 / B: / THR / 146 / [ / O / ]
364 / A: / VAL / 158 / [ / CB / ] / 3.75 / B: / THR / 146 / [ / O / ]
365 / A: / PHE / 159 / [ / CA / ] / 3.68 / B: / SER / 144 / [ / O / ]
366 / A: / PHE / 159 / [ / C / ] / 3.72 / B: / SER / 144 / [ / O / ]
367 / A: / ARG / 160 / [ / N / ] / 2.85 / B: / SER / 144 / [ / O / ]
368 / A: / ARG / 160 / [ / N / ] / 3.91 / B: / SER / 144 / [ / C / ]
369 / A: / ARG / 160 / [ / N / ] / 3.95 / B: / ASN / 145 / [ / CA / ]
370 / A: / ARG / 160 / [ / CA / ] / 3.75 / B: / SER / 144 / [ / O / ]
371 / A: / ARG / 160 / [ / CB / ] / 3.65 / B: / SER / 144 / [ / O / ]
372 / A: / ARG / 160 / [ / CG / ] / 3.85 / B: / ASN / 145 / [ / CA / ]
373 / A: / ARG / 160 / [ / CG / ] / 3.96 / B: / ASN / 145 / [ / C / ]
374 / A: / ARG / 160 / [ / CG / ] / 3.84 / B: / THR / 146 / [ / OG1 / ]
375 / A: / ARG / 160 / [ / CD / ] / 3.81 / B: / THR / 146 / [ / OG1 / ]
376 / A: / ARG / 160 / [ / NE / ] / 3.58 / B: / THR / 146 / [ / CB / ]
377 / A: / ARG / 160 / [ / NE / ] / 2.69 / B: / THR / 146 / [ / OG1 / ]
378 / A: / ARG / 160 / [ / CZ / ] / 3.28 / B: / THR / 146 / [ / OG1 / ]
379 / A: / ARG / 160 / [ / NH2 / ] / 3.68 / B: / THR / 146 / [ / CB / ]
380 / A: / ARG / 160 / [ / NH2 / ] / 3.95 / B: / THR / 146 / [ / CG2 / ]
381 / A: / ARG / 160 / [ / NH2 / ] / 3.04 / B: / THR / 146 / [ / OG1 / ]
382 / A: / ARG / 161 / [ / NH1 / ] / 3.63 / B: / GLN / 143 / [ / NE2 / ]
383 / A: / ARG / 186 / [ / CD / ] / 3.91 / B: / GLN / 101 / [ / NE2 / ]
384 / A: / ARG / 186 / [ / NE / ] / 4.00 / B: / GLN / 101 / [ / CD / ]
385 / A: / ARG / 186 / [ / NE / ] / 3.44 / B: / GLN / 101 / [ / NE2 / ]
386 / A: / ARG / 186 / [ / CZ / ] / 3.67 / B: / GLN / 101 / [ / CD / ]
387 / A: / ARG / 186 / [ / CZ / ] / 3.27 / B: / GLN / 101 / [ / NE2 / ]
388 / A: / ARG / 186 / [ / NH1 / ] / 3.52 / B: / GLN / 101 / [ / NE2 / ]
389 / A: / ARG / 186 / [ / NH2 / ] / 3.89 / B: / GLN / 101 / [ / CG / ]
390 / A: / ARG / 186 / [ / NH2 / ] / 3.54 / B: / GLN / 101 / [ / CD / ]
391 / A: / ARG / 186 / [ / NH2 / ] / 3.86 / B: / GLN / 101 / [ / OE1 / ]
392 / A: / ARG / 186 / [ / NH2 / ] / 3.63 / B: / GLN / 101 / [ / NE2 / ]
393 / A: / GLU / 341 / [ / CD / ] / 3.98 / B: / SER / 144 / [ / CB / ]
394 / A: / GLU / 341 / [ / CD / ] / 3.25 / B: / SER / 144 / [ / OG / ]
395 / A: / GLU / 341 / [ / OE1 / ] / 3.04 / B: / SER / 144 / [ / CB / ]
396 / A: / GLU / 341 / [ / OE1 / ] / 2.64 / B: / SER / 144 / [ / OG / ]
397 / A: / GLU / 341 / [ / OE2 / ] / 3.81 / B: / GLU / 142 / [ / OE1 / ]
398 / A: / GLU / 341 / [ / OE2 / ] / 3.76 / B: / ASN / 145 / [ / ND2 / ]
399 / A: / GLU / 341 / [ / OE2 / ] / 3.18 / B: / SER / 144 / [ / OG / ]

Supplementary Table S2 List of interactions (≤ 4 Å) between maltose (MAL) and surrounding residues. Interactions of the catalytic D322 with O1 oxygen atom of maltose are highlighted in green.

No. / Maltose / Dist. [Å] / Chain B
1 / C: / MAL / 1 / [ / C6' / ] / 3.96 / B: / TYR / 433 / [ / CZ / ]
2 / C: / MAL / 1 / [ / C6' / ] / 3.56 / B: / TYR / 433 / [ / OH / ]
3 / C: / MAL / 1 / [ / C5' / ] / 3.91 / B: / TYR / 433 / [ / OH / ]
4 / C: / MAL / 1 / [ / C5' / ] / 3.72 / B: / HIS / 324 / [ / NE2 / ]
5 / C: / MAL / 1 / [ / O5' / ] / 3.43 / B: / TYR / 433 / [ / CE1 / ]
6 / C: / MAL / 1 / [ / O5' / ] / 3.33 / B: / TYR / 433 / [ / CZ / ]
7 / C: / MAL / 1 / [ / O5' / ] / 3.12 / B: / TYR / 433 / [ / OH / ]
8 / C: / MAL / 1 / [ / C1' / ] / 3.57 / B: / TYR / 433 / [ / CE1 / ]
9 / C: / MAL / 1 / [ / O1' / ] / 3.71 / B: / HIS / 324 / [ / NE2 / ]
10 / C: / MAL / 1 / [ / O1' / ] / 3.07 / B: / ASP / 322 / [ / OD2 / ]
11 / C: / MAL / 1 / [ / O1' / ] / 3.88 / B: / ASP / 322 / [ / CG / ]
12 / C: / MAL / 1 / [ / O1' / ] / 3.88 / B: / ASP / 322 / [ / OD1 / ]
13 / C: / MAL / 1 / [ / C4' / ] / 3.93 / B: / HIS / 324 / [ / NE2 / ]
14 / C: / MAL / 1 / [ / C3' / ] / 3.74 / B: / HIS / 324 / [ / NE2 / ]
15 / C: / MAL / 1 / [ / C3' / ] / 3.86 / B: / HIS / 324 / [ / CD2 / ]
16 / C: / MAL / 1 / [ / C3' / ] / 3.85 / B: / ASP / 322 / [ / OD1 / ]
17 / C: / MAL / 1 / [ / O3' / ] / 3.22 / B: / LYS / 426 / [ / NZ / ]
18 / C: / MAL / 1 / [ / O3' / ] / 3.59 / B: / TYR / 429 / [ / CE2 / ]
19 / C: / MAL / 1 / [ / C2' / ] / 3.89 / B: / TYR / 433 / [ / CD1 / ]
20 / C: / MAL / 1 / [ / C2' / ] / 3.56 / B: / TYR / 429 / [ / OH / ]
21 / C: / MAL / 1 / [ / C2' / ] / 3.74 / B: / TYR / 433 / [ / CE1 / ]
22 / C: / MAL / 1 / [ / C2' / ] / 3.84 / B: / ASP / 322 / [ / OD1 / ]
23 / C: / MAL / 1 / [ / O2' / ] / 3.59 / B: / TYR / 429 / [ / CE2 / ]
24 / C: / MAL / 1 / [ / O2' / ] / 3.71 / B: / TYR / 429 / [ / CZ / ]
25 / C: / MAL / 1 / [ / O2' / ] / 2.97 / B: / TYR / 429 / [ / OH / ]
26 / C: / MAL / 1 / [ / O2' / ] / 3.31 / B: / ASP / 322 / [ / OD2 / ]
27 / C: / MAL / 1 / [ / O2' / ] / 3.94 / B: / PRO / 344 / [ / CD / ]
28 / C: / MAL / 1 / [ / O2' / ] / 3.33 / B: / ASP / 322 / [ / CG / ]
29 / C: / MAL / 1 / [ / O2' / ] / 2.72 / B: / ASP / 322 / [ / OD1 / ]
30 / C: / MAL / 1 / [ / O1 / ] / 3.95 / B: / HIS / 324 / [ / CE1 / ]
31 / C: / MAL / 1 / [ / O1 / ] / 3.57 / B: / HIS / 324 / [ / NE2 / ]
32 / C: / MAL / 1 / [ / O1 / ] / 3.67 / B: / HIS / 324 / [ / CD2 / ]
33 / C: / MAL / 1 / [ / O5 / ] / 3.32 / B: / ARG / 438 / [ / NH2 / ]
34 / C: / MAL / 1 / [ / C6 / ] / 3.99 / B: / TRP / 222 / [ / CE3 / ]
35 / C: / MAL / 1 / [ / C6 / ] / 3.72 / B: / TRP / 222 / [ / CZ3 / ]
36 / C: / MAL / 1 / [ / C6 / ] / 3.79 / B: / TRP / 222 / [ / CH2 / ]
37 / C: / MAL / 1 / [ / O6 / ] / 3.55 / B: / ARG / 438 / [ / CZ / ]
38 / C: / MAL / 1 / [ / O6 / ] / 3.02 / B: / ARG / 438 / [ / NH2 / ]
39 / C: / MAL / 1 / [ / O6 / ] / 3.29 / B: / ARG / 438 / [ / NH1 / ]
40 / C: / MAL / 1 / [ / C4 / ] / 3.87 / B: / SER / 367 / [ / OG / ]
41 / C: / MAL / 1 / [ / O4 / ] / 3.95 / B: / SER / 367 / [ / CA / ]
42 / C: / MAL / 1 / [ / O4 / ] / 3.64 / B: / SER / 367 / [ / CB / ]
43 / C: / MAL / 1 / [ / O4 / ] / 3.79 / B: / TRP / 222 / [ / CE3 / ]
44 / C: / MAL / 1 / [ / O4 / ] / 3.58 / B: / TRP / 222 / [ / CZ3 / ]
45 / C: / MAL / 1 / [ / O4 / ] / 2.74 / B: / SER / 367 / [ / OG / ]
46 / C: / MAL / 1 / [ / O4 / ] / 3.63 / B: / HIS / 324 / [ / CB / ]
47 / C: / MAL / 1 / [ / O4 / ] / 3.72 / B: / HIS / 324 / [ / CG / ]
48 / C: / MAL / 1 / [ / O4 / ] / 3.49 / B: / HIS / 324 / [ / ND1 / ]
49 / C: / MAL / 1 / [ / C3 / ] / 3.67 / B: / HIS / 324 / [ / CB / ]
50 / C: / MAL / 1 / [ / C3 / ] / 3.71 / B: / HIS / 324 / [ / CG / ]
51 / C: / MAL / 1 / [ / O3 / ] / 3.87 / B: / TYR / 370 / [ / CD2 / ]
52 / C: / MAL / 1 / [ / O3 / ] / 3.46 / B: / SER / 367 / [ / N / ]
53 / C: / MAL / 1 / [ / O3 / ] / 3.15 / B: / SER / 367 / [ / CA / ]
54 / C: / MAL / 1 / [ / O3 / ] / 3.44 / B: / SER / 367 / [ / CB / ]
55 / C: / MAL / 1 / [ / O3 / ] / 3.99 / B: / GLU / 430 / [ / OE2 / ]
56 / C: / MAL / 1 / [ / O3 / ] / 3.67 / B: / SER / 367 / [ / OG / ]
57 / C: / MAL / 1 / [ / O3 / ] / 3.59 / B: / HIS / 324 / [ / CB / ]
58 / C: / MAL / 1 / [ / C2 / ] / 3.93 / B: / TYR / 370 / [ / CE1 / ]
59 / C: / MAL / 1 / [ / C2 / ] / 3.89 / B: / TYR / 370 / [ / CE2 / ]
60 / C: / MAL / 1 / [ / C2 / ] / 3.83 / B: / TYR / 370 / [ / CZ / ]
61 / C: / MAL / 1 / [ / O2 / ] / 3.94 / B: / GLU / 430 / [ / CD / ]
62 / C: / MAL / 1 / [ / O2 / ] / 3.7 / B: / GLU / 430 / [ / CG / ]
63 / C: / MAL / 1 / [ / O2 / ] / 3.29 / B: / GLU / 430 / [ / OE2 / ]
64 / C: / MAL / 1 / [ / O2 / ] / 3.72 / B: / LYS / 426 / [ / CE / ]
65 / C: / MAL / 1 / [ / O2 / ] / 3.95 / B: / LYS / 426 / [ / NZ / ]

Supplementary Table S3List of primers used in molecular cloning and mutagenesis. Restriction sites are underlined.

Forward primer / Reverse primer
Mak / 5’-cctgGGATCCATGACTCGGTCGGACACGCT-3’ / 5’-gccgCTCGAGTCAGCTAGCGGTCAGGCGGG-3’
W222A / 5’-GAAGGCGCAGCGATGGCCACCGCCAGCGTG-3’ / 5’-GGCCGCGTTCGCCTCATACTCGGTCACCAT-3’
H324A / 5’-GACCTGGCATTGGGACAGGTGCTGCGTACC -3’ / 5’-ACCGTGCACCCGCTGGACCGTGATTGCCTC -3’
Y433A / 5’-ACCGGCGCAGAGACACGGCACCGGCCGGGT-3’ / 5’-CTCATAAACCGCCTTGTCGAGTTCGTAGGC -3’
Y370A / 5’-TTCGAGGCAGCCGCTTACGGGCCGCTGGTGG-3’ / 5’-CGATCGCAACACACCGGCCACGTCGCGCA-3’
E430A / 5’-GTTTATGCAACCGGCTATGAGACACGGCACC -3’ / 5’-CGCCTTGTCGAGTTCGTAGGCGCCCAACA-3’
Y429A / 5’-GCGGTTGCAGAGACCGGCTATGAGACACGG-3’ / 5’-CTTGTCGAGTTCGTAGGCGCCCAACAGCAG-3’
D322A / 5’-CACGGTGCACTGCACTTGGGACAGGTGCTGC-3’ / 5’-CACCCGCTGGACCGTGATTGCCTCCGCCGC-3’
K157A / 5’-ATCCTCGCAGTGTTCCGCCGGGTAAGCAGCG-3’ / 5’-AGCCCGCCGATCGAAGATCACACTGGTGTT-3’
D169A / 5’-CCCGCAATAGAGCTGAACCGCGTGCTTACCC-3’ / 5’-GTTGATCCCGCTGCTTACCCGGCGGAACAC-3’
N173A / 5’-GAGCTGGCACGCGTGCTTACCCGTGCCGGTA-3’ / 5’-TATGTCGGGGTTGATCCCGCTGCTTACCCG-3’
T212A / 5’-ATGGTGGCAGAGTATGAGGCGAACGCGGCC-3’ / 5’-GCCCAGGGCGTACGCCAGAGCATCGGTTG-3’
Y214A / 5’-ACCGAGGCAGAGGCGAACGCGGCCGAAGG-3’ / 5’-CACCATGCCCAGGGCGTACGCCAGAGCATC-3’
E220A / 5’-GCGGCCGCAGGCTGGGCGATGGCCACCGCC-3’ / 5’-GTTCGCCTCATACTCGGTCACCATGCCCAG-3’
Q327A / 5’-TTGGGAGCAGTGCTGCGTACCCCGGAAAGCT-3’ / 5’-GTGCAGGTCACCGTGCACCCGCTGGACCGT-3’
D339A / 5’-TTGATCGCATTTGAAGGCGAGCCGGGCCAGC-3’ / 5’-CAGCCAGCTTTCCGGGGTACGCAGCACCTG-3’
E341A / 5’-GACTTTGCAGGCGAGCCGGGCCAGCCGCTG-3’ / 5’-GATCAACAGCCAGCTTTCCGGGGTACGCAG-3’
E142A / 5’-GACGCCGCACAGAGCAACACCAGTGTGATCT-3’ / 5’-ACATACCCGCGGCTGCGCGGCAAAGGGCA-3’
Q143A / 5’-GCCGAAGCAAGCAACACCAGTGTGATCTTCG-3’ / 5’-GTCACATACCCGCGGCTGCGCGGCAAAGG-3’
S144A / 5’-GAACAGGCAAACACCAGTGTGATCTTCGATC-3’ / 5’-GGCGTCACATACCCGCGGCTGCGCGGCAA-3’
I155A / 5’-CGGGCTGCACTCAAGGTGTTCCGCCGGGTAA-3’ / 5’-CCGATCGAAGATCACACTGGTGTTGCTCTG-3’
L329A / 5’-CAGGTGGCACGTACCCCGGAAAGCTGGCTG-3’ / 5’-TCCCAAGTGCAGGTCACCGTGCACCCGCTG-3’
I338A / 5’-CTGTTGGCAGACTTTGAAGGCGAGCCGGGC-3’ / 5’-CCAGCTTTCCGGGGTACGCAGCACCTGTC-3’
R438A / 5’-CGGCACGCACCGGGTTGGCTTCCGATTCCGC-3’ / 5’-TGTCTCATAGCCGGTCTCATAAACCGCCTT-3’
S367A / 5’-TTGCGAGCATTCGAGTACGCCGCTTACGGGC-3’ / 5’-CACACCGGCCACGTCGCGCAGCGGCGAAT-3’

Supplementary Table S4 Enzyme activity data of MtbMaK wild type and mutants. These data were used for making Figure 3f and 4c of the main text.Error represent s.d. (n=3).

Enzyme / Enzyme activity (μmol.min-1.mg-1) / Relative activity (%)
Wild type / 12.36±0.28 / 100
W222A / 1.57±0.02 / 12.71
D322A / 0.18±0.00 / 1.44
H324A / 0.52±0.27 / 4.18
S367A / 0.71±0.04 / 5.78
Y370A / 0.68±0.01 / 5.52
Y429A / 0.62±0.20 / 5.02
E430A / 1.12±0.05 / 9.09
Y433A / 0.41±0.16 / 3.31
R438A / 1.07±0.01 / 8.63
I155A / 3.38±0.15 / 27.35
I338A / 4.43±0.14 / 35.88
Y214A / 0.53±0.02 / 4.31
L329A / 0.92±0.04 / 7.42
N173A / 10.99±0.50 / 88.92
T212A / 11.83±0.42 / 95.73
E220A / 4.84±0.30 / 39.17
K157A / 1.74±0.02 / 14.11
D169A / 1.63±0.02 / 13.15
Q327A / 0.24±0.04 / 1.94
D339A / 0.36±0.05 / 2.92
E341A / 0.35±0.06 / 2.80
E142A / 1.86±0.02 / 15.02
Q143A / 2.88±0.15 / 23.30
S144A / 0.45±0.02 / 3.62

1