Supplemental Material, Table 1. Sequence identity and similarity of retroviral proteases.

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PRHIV-1 HIV-2 EIAV AMV MPMV MMTV MMLV HTLV-1 BLV WDSV HFV

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HIV-1 ------

HIV-247/63 ------

65/80

EIAV31/40 36/46------

70/75 55/75

AMV31/41 26/40 27/45 ------

50/55 50/55 45/60

MPMV 24/39 22/36 20/37 25/41 ------

50/75 45/65 40/75 40/70

MMTV22/32 20/31 22/37 24/38 46/57 ------

45/65 40/55 40/65 35/60 55/65

MMLV24/41 24/36 24/35 16/33 21/32 16/30 ------

40/50 35/45 40/50 25/40 35/50 40/55

HTLV-134/42 26/40 29/42 24/37 20/33 18/27 22/29 ------

50/55 45/50 50/55 40/50 30/55 40/55 45/55

BLV28/45 22/39 25/38 22/34 17/30 15/27 27/36 32/43 ------

45/55 40/50 45/55 35/50 25/55 30/50 55/60 75/85

WDSV15/26 19/32 22/29 17/28 19/28 18/32 22/34 19/29 13/21 -- --

35/35 40/40 30/35 35/40 35/45 30/40 25/35 30/40 30/35

HFV23/30 16/30 15/25 22/28 12/20 10/19 15/21 17/26 18/27 14/22 --

35/35 25/40 25/30 30/30 25/40 20/30 15/20 15/25 15/20 25/40

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Sequence identity and similarity of full retroviral PR sequences as well as of residues involved in substrate binding (numbers in bold) are given as percentage identity/similarity.

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Supplemental material, Table 2. Relative activities obtained on Val-Ser-Gln-Asn-XaaPro-Ile-Val-Gln peptides for retroviral proteases.

______

Lentivirus1Alpha- BetaretrovirusGamma- Deltaretrovirus Epsilon-

retrovirusretrovirus retrovirus

____

Protease of

P1 residueHIV-1HIV-2EIAVAMVMPMVMMTVMMLVHTLV-1BLV WDSV ______

Gly00000000nd20

Ala0.01000.0100000.270

Val0000.0200000.130

Leu0.210.250.090.210.030.010.91nd100.30

Phe1.71.31.41.62.11.38.207.61.8

Tyr1111111011

Trp0.01nd00.0200000.180

Cys <0.01nd0.69<0.010nd000.86nd

Met0.230.400.150.170.020.010.7006.80.51

Ser0000000000

Asp00000.0200nd00

Lys0000000000

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1Classification is based on the latest report of the International Committee on Taxonomy of Viruses (ICTV), as detailed at the ICTV database ( were expressed relative to that obtained with the Val-Ser-Gln-Asn-TyrPro-Ile-Val-Gln substrate, except for HTLV-1 and HFV PRs. 2nd, not determined.The best two values are highlighted in bold.It should be noted, that none of the substrates was cleaved by HFV PR (data not shown).

Supplemental Material, Table 3. Relative activities obtained on Val-Ser-Xaa-Asn-TyrPro-Ile-Val-Gln peptides for retroviral proteases.

______

Lentivirus1Alpha- BetaretrovirusGamma- Deltaretrovirus Epsilon-

retrovirusretrovirus retrovirus

Protease of

P3 residueHIV-1HIV-2EIAVAMVMPMVMMTVMMLVHTLV-1BLV WDSV

______

Gly0.180.230.210.030.5900.1800.230.67

Ala0.340.300.140.420.5100.5501.80.42

Val0.710.640.010.500.461.40.0200.230.32

Leu0.090.420.012.01.04.80.1000.280.35

Phe0.220.130.232.41.37.50.7700.050.11

Asn0.500.320.080.050.431.10.8100.260.11

Gln1111111011

Asp0.010.1000.010.810.060.0100.220

Lys0.120.160.010.070.070.510.4001.70.34

______

1Classification is based on the latest report of the International Committee on Taxonomy of Viruses (ICTV), as detailed at the ICTV database ( were expressed relative to that obtained with the Val-Ser-Gln-Asn-TyrPro-Ile-Val-Gln substrate, except for HFV PR for which only one peptide was cleaved. The best two values are highlighted in bold.It should be noted, that in case of HFV, only the P3 Val-containing peptide was hydrolyzed, therefore it was not possible to calculate the relative activity value.

Supplemental Material, Table 4. Relative activities obtained on Val-Xaa-Gln-Asn-TyrPro-Ile-Val-Gln peptides for retroviral proteases.

______

Lentivirus1Alpha- BetaretrovirusGamma- Deltaretrovirus Epsilon-

retrovirusretrovirus retrovirus

Protease of

P4 residueHIV-1HIV-2EIAVAMVMPMVMMTVMMLVHTLV-1BLV WDSV

______

Gly15270.090.020.110.081600.030.10

Ala6.0100.860.240.570.561401.21.9

Val1111111111

Ile0.601.00.862.11.72.26.70.920.071.4

Leu0.402.01.11.22.40.643.3nd01.0

Phe0.600.330.931.13.14.48.00.7701.0

Met0.807.30.460.641.30.72180.3801.0

Ser20330.710.110.290.403301.10.43

Thr5.4190.240.530.43nd2601.00.48

Pro2.84.01.40.680.34nd151.71.61.3

Asn3.4nd0.490.150.340.2310nd0.030.43

Asp3.4210.490.090.130.1423000.03

Lys1.21.00.180.322.107.300.011.0

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1Classification is based on the latest report of the International Committee on Taxonomy of Viruses (ICTV), as detailed at the ICTV database ( Relative activities were expressed as activity relative to that obtained with the Val-Val-Gln-Asn-TyrPro-Ile-Val-Gln substrate instead of those obtained with the original P4-Ser-containing one to allow the inclusion of the HTLV-1 PR data.The best two values are highlighted in bold. None of the substrates was cleaved by HFV PR (data not shown).

1