Supporting Information
For
Alumina Nanocomposites-A Comparative Approach Highlighting the Improved Characteristics of Nanocomposites for PhosphopeptidesEnrichment
Muhammad Najam-ul-Haq*, Fahmida Jabeen, Batool Fatima, Muhammad Naeem Ashiq, Dilshad Hussain
Division of Analytical Chemistry, Institute of Chemical Sciences, Bahauddin Zakariya University, Multan 60800, Pakistan
Chemicals and Materials
Aluminum oxide (particle size <50 nm), lanthanum oxide (particle size <100 nm, 99%), ceria (particle size <50 nm, 99.95%) , titania (particle size 21 nm, 99.5%) , zirconia (particle size <100 nm) nanoparticles, aluminum chloride (99.9%), zirconium IV chloride (99%), titanium IV chloride (98%), lanthanum chloride hydrate (99.9%), cerium III chloride heptahydrate (99.9%), ammonium hydroxide solution, bovine β-casein, trifluoroacetic acid (TFA), acetonitrile (ACN), ammonium bicarbonate (NH4HCO3),2,5-Dihydroxybenzoic acid (2,5-DHB), dithiothreitol (DTT) and iodoacetamide (IAA) were purchased from Sigma Aldrich. Trypsin was obtained from Promega whereas MilliQ water was provided by the laboratory.
Fig. S1:EDAX spectra showing the elemental composition of (a) alumina-ceria and (b) alumina-lanthanum oxide nanocomposites.
Fig. S2: EDX spectra of (a) alumina-titania and (b) alumina-zirconia nanocomposites.
Fig. S3:MALDI-MS spectra for sensitivity assessment of 10 femtomole β-casein digest. (A) Nanocomposites of (a) alumina-ceria (b) alumina-zirconia (c) alumina-titania and (d) lanthanum oxide. (B) Nanoparticles of individual metal oxides (a) ceria (b) zirconia (c) titania and (d) lanthanum oxide.
Fig. S4:The serum analysis using four nanocomposites. (a) Percentages of identified phosphoproteins in serum by the nanocomposites and (b) percentage of absent phosphoproteins in serum by nanocomposites.
Table S1: Comparative data for four nanocomposites used to perform enrichment from tryptic serum digest. The phosphoproteins are characteristic of enriched phosphopeptides identified in eluted fractions. The symbols showing presence and absence of certain phosphoproteins, “$” and ‘’0’’ respectively.
Sr.No / Identified Protein / Alumina-zirconia / Alumina-titania / Alumina-
ceria / Alumina-
lanthanum oxide
1 / Peripherin / $ / $ / $ / $
2 / Purine nucleoside phosphorylase / $ / $ / 0 / 0
3 / Ellis-van Creveld syndrome protein / $ / $ / 0 / $
4 / Ras-related protein Rap-1A / $ / $ / 0 / $
5 / 3'(2'),5'-bisphosphate nucleotidase 1 / $ / $ / 0 / 0
6 / DnaJ homolog subfamily B member 9 / $ / $ / 0 / $
7 / E3 ubiquitin-protein ligase RING1 / $ / $ / 0 / $
8 / Carbonic anhydrase 3 / $ / $ / $ / 0
9 / E3 ubiquitin-protein ligase RNF14 / $ / $ / 0 / $
10 / PRAME family member 2 / $ / $ / 0 / 0
11 / Dehydrogenase/reductase SDR family member 4 / $ / $ / 0 / $
12 / Protein FAM69A / $ / 0 / 0 / 0
13 / Putative serine protease 46 / $ / $ / 0 / $
14 / Ankyrin repeat and SOCS box protein 7 / $ / $ / $ / 0
15 / Kinesin-like protein KIF1B / $ / $ / $ / $
16 / Solute carrier family 25 member 38 / $ / $ / 0 / 0
17 / Collagen alpha-1(XXV) chain / $ / - / 0 / $
18 / tRNA (guanine-N(7)-)-methyltransferase non-catalytic subunit WDR4 / $ / $ / 0 / 0
19 / Putative zinc finger protein 75C / $ / $ / 0 / $
20 / Polymerase delta-interacting protein 3 / 0 / $ / 0 / $
21 / Centromere protein P / $ / $ / 0 / $
22 / Replication factor C subunit 2 / $ / $ / 0 / $
23 / Developmentally-regulated GTP-binding protein 2 / 0 / $ / 0 / 0
24 / N-acyl-phosphatidylethanolamine-hydrolyzing phospholipase D / $ / $ / 0 / $
25 / Zinc finger protein 793 / $ / $ / 0 / 0
26 / Protein RTF2 homolog / $ / $ / 0 / $
27 / Putative hexokinase HKDC1 / 0 / $ / 0 / $
28 / Src kinase-associated phosphoprotein 2 / $ / $ / $ / $
29 / Signal peptidase complex subunit 3 / 0 / $ / 0 / $
30 / Prostate and testis expressed protein 4 / 0 / $ / 0 / $
31 / Calcium-binding protein 7 / $ / 0 / 0 / $
32 / Caspase-4 / 0 / $ / 0 / $
33 / Beta-defensin 132 / 0 / $ / 0 / $
34 / Keratin-associated protein 13-3 / 0 / 0 / 0 / $
35 / Mitochondrial thiamine pyrophosphate carrier / 0 / 0 / 0 / $
36 / Sperm-associated antigen 11B / 0 / 0 / $ / 0
37 / A-kinase anchor protein 4 / 0 / 0 / $ / 0
38 / Heat shock transcription factor, y-linked / 0 / 0 / 0 / $
39 / Sphingosine 1-phosphate receptor 2 / 0 / 0 / $ / $
40 / Cancer-related nucleoside-triphosphatase / 0 / 0 / 0 / $
41 / Deoxycytidine kinase / 0 / 0 / $ / 0
42 / Zinc finger protein ZFPM2 / 0 / 0 / $ / $
43 / Bromodomain-containing protein 1 / 0 / 0 / $ / 0
44 / Glycogen phosphorylase / 0 / 0 / 0 / $
45 / Myosin-IIIb / 0 / 0 / 0 / $
46 / Selenium-binding protein 1 / 0 / 0 / 0 / $
47 / Tyrosine-protein kinase JAK1 / 0 / 0 / $ / $
48 / Breast cancer type 2 susceptibility protein / 0 / 0 / 0 / $
49 / FERM domain-containing protein 6 / 0 / 0 / $ / 0
50 / Hydroxysteroid dehydrogenase-like protein 2 / 0 / 0 / 0 / $
51 / 14-3-3 protein epsilon / 0 / 0 / $ / $
52 / Insulinoma-associated protein 2 / 0 / 0 / 0 / $
53 / Growth factor receptor-bound protein 7 / 0 / 0 / 0 / $
54 / Coiled-coil domain-containing protein 171 / 0 / 0 / $ / $
55 / Erlin-1 / 0 / 0 / 0 / $
56 / Ellis-van Creveld syndrome protein / 0 / 0 / $ / $
57 / Purine nucleoside phosphorylase / 0 / 0 / 0 / $
58 / Calcium-activated potassium channel subunit alpha-1 / 0 / 0 / $ / $
59 / Probable cysteine--tRNA ligase, mitochondrial / 0 / 0 / 0 / $
60 / NADH dehydrogenase [ubiquinone] flavoprotein 1, mitochondrial / 0 / 0 / 0 / $
61 / Serine protease 23 / 0 / 0 / $ / $
62 / Transcription factor PU.1 O / 0 / 0 / 0 / $
63 / Sperm protein associated with the nucleus on the X chromosome N4 / 0 / 0 / $ / $
64 / Protein naked cuticle homolog 1 / 0 / 0 / 0 / $
65 / Ubiquitin carboxyl-terminal hydrolase 8 / 0 / 0 / $ / $
66 / 28S ribosomal protein S2, mitochondrial / 0 / 0 / 0 / $
67 / Replication factor C subunit 5 / 0 / 0 / 0 / $
68 / GDNF-inducible zinc finger protein 1 / 0 / 0 / $ / 0
69 / UBX domain-containing protein 11 / 0 / 0 / 0 / $
70 / Putative deoxyribonuclease TATDN1 / 0 / 0 / $ / 0
71 / WD repeat-containing protein 78 / 0 / 0 / 0 / $
72 / Mitochondrial import inner membrane translocase subunit Tim13 / 0 / 0 / $ / 0
73 / Hydroxysteroid dehydrogenase-like protein 2 / 0 / 0 / 0 / $
74 / NADH dehydrogenase [ubiquinone] flavoprotein 1, mitochondrial / 0 / 0 / $ / $
75 / Cyclin-H / 0 / 0 / $ / 0
76 / Putative zinc finger protein 75C / 0 / 0 / $ / $
77 / ATP-dependent RNA helicase DHX8 / 0 / 0 / 0 / $
78 / E3 ubiquitin-protein ligase RING1 / 0 / 0 / $ / 0
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