nsP3

549 residues (nt 4101-5747) 60288 dalton

(In theory, nsP3 produced by cleavage has a 6-residue carboxyl extension compared to nsP3 produced by terminatation at the opal codon. The sequence below ends at the opal codon).

Sequence (see also comparison with other alphaviruses)

             10         20         30         40         50
  0  APSYRTKREN IADCQEEAVV NAANPLGRPG EGVCRAIYKR WPTSFTDSAT
 50  ETGTARMTVC LGKKVIHAVG PDFRKHPEAE ALKLLQNAYH AVADLVNEHN
100  IKSVAIPLLS TGIYAAGKDR LEVSLNCLTT ALDRTDADVT IYCLDKKWKE
150  RIDAALQLKE SVTELKDEDM EIDDELVWIH PDSCLKGRKG FSTTKGKLYS
200  YFEGTKFHQA AKDMAEIKVL FPNDQESNEQ LCAYILGETM EAIREKCPVD
250  HNPSSSPPKT LPCLCMYAMT PERVHRLRSN NVKEVTVCSS TPLPKHKIKN
300  VQKVQCTKVV LFNPHTPAFV PARKYIEVPE QPTAPPAQAE EAPEVVATPS
350  PSTADNTSLD VTDISLDMDD SSEGSLFSSF SGSDNSITSM DSWSSGPSSL
400  EIVDRRQVVV ADVHAVQEPA PIPPPRLKKM ARLAAARKEP TPPASNSSES
450  LHLSFGGVSM SLGSIFDGET ARQAAVQPLA TGPTDVPMSF GSFSDGEIDE
500  LSRRVTESEP VLFGSFEPGE VNSIISSRSA VSFPLRKQRR RRRSRRTEY

Composition (mole %)

Ala 48 (8.7 %) Asx  0 ( 0.0 %) Cys 12 (2.2 %) Asp 32 (5.8 %) Glu 42 (7.7 %)
Phe 17 (3.1 %) Gly 25 ( 4.6 %) His 12 (2.2 %) Ile 23 (4.2 %) Lys 33 (6.0 %)
Leu 40 (7.3 %) Met 11 ( 2.0 %) Asn 18 (3.3 %) Pro 42 (7.7 %) Gln 15 (2.7 %)
Arg 33 (6.0 %) Ser 55 (10.0 %) Thr 35 (6.4 %) Val 41 (7.5 %) Trp  4 (0.7 %)
Tyr 11 (2.0 %) Glx  0 ( 0.0 %) xxx  0 (0.0 %)

Titration of nsP3 (number of residues protonated)

(-) (+) net pH Cys Asp Glu Tyr COOH charge His Lys Arg NH2 charge charge
1.0 12.00 31.96 41.98 11.00 1.00 -0.06 12.00 33.00 33.00 1.00 79.00 78.94
1.5 12.00 31.88 41.93 11.00 0.99 -0.20 12.00 33.00 33.00 1.00 79.00 78.80
2.0 12.00 31.61 41.77 11.00 0.97 -0.65 12.00 33.00 33.00 1.00 79.00 78.35
2.5 12.00 30.80 41.27 11.00 0.92 -2.01 12.00 33.00 33.00 1.00 79.00 76.99
3.0 12.00 28.50 39.76 11.00 0.78 -5.96 12.00 33.00 33.00 1.00 79.00 73.04
3.5 12.00 23.04 35.66 11.00 0.53 -15.77 11.99 33.00 33.00 1.00 78.99 63.22
4.0 12.00 14.35 26.88 11.00 0.27 -33.50 11.96 33.00 33.00 1.00 78.96 45.46
4.5 12.00 6.54 15.12 11.00 0.10 -53.24 11.88 33.00 33.00 1.00 78.88 25.64
5.0 11.99 2.41 6.34 11.00 0.04 -66.22 11.63 33.00 33.00 1.00 78.63 12.41
5.5 11.98 0.80 2.24 11.00 0.01 -71.97 10.91 33.00 33.00 1.00 77.91 5.94
6.0 11.94 0.26 0.73 11.00 0.00 -74.07 9.12 33.00 33.00 1.00 76.12 2.05
6.5 11.81 0.08 0.23 11.00 0.00 -74.88 6.00 33.00 33.00 0.99 72.99 -1.89
7.0 11.43 0.03 0.07 11.00 0.00 -75.47 2.88 32.99 33.00 0.97 69.84 -5.63
7.5 10.36 0.01 0.02 11.00 0.00 -76.61 1.09 32.98 33.00 0.92 67.99 -8.62
8.0 7.99 0.00 0.01 10.99 0.00 -79.01 0.37 32.95 33.00 0.78 67.10 -11.91
8.5 4.64 0.00 0.00 10.96 0.00 -82.40 0.12 32.83 33.00 0.53 66.48 -15.92
9.0 2.00 0.00 0.00 10.88 0.00 -85.12 0.04 32.47 32.99 0.27 65.77 -19.35
9.5 0.71 0.00 0.00 10.62 0.00 -86.67 0.01 31.39 32.97 0.10 64.47 -22.20
10.0 0.23 0.00 0.00 9.89 0.00 -87.88 0.00 28.39 32.90 0.04 61.33 -26.55
10.5 0.08 0.00 0.00 8.12 0.00 -89.80 0.00 21.81 32.67 0.01 54.49 -35.31
11.0 0.02 0.00 0.00 5.18 0.00 -92.80 0.00 12.59 31.99 0.00 44.58 -48.22
11.5 0.01 0.00 0.00 2.42 0.00 -95.57 0.00 5.38 30.00 0.00 35.38 -60.19
12.0 0.00 0.00 0.00 0.90 0.00 -97.10 0.00 1.92 25.07 0.00 26.99 -70.11
12.5 0.00 0.00 0.00 0.30 0.00 -97.70 0.00 0.63 16.50 0.00 17.13 -80.57
13.0 0.00 0.00 0.00 0.10 0.00 -97.90 0.00 0.20 7.93 0.00 8.13 -89.77

Isoelectric point

                                          (-)                                (+)    net 
 pI    Cys    Asp    Glu    Tyr   COOH   charge  His    Lys    Arg    NH2  charge  charge
6.26  11.89   0.14   0.40  11.00  0.00  -74.57   7.58  33.00  33.00  0.99  74.57    0.00

The conserved domain

326 residues (4101-5078) 36472 dalton

Sequence

            10         20         30         40         50
  0  APSYRTKREN IADCQEEAVV NAANPLGRPG EGVCRAIYKR WPTSFTDSAT
 50  ETGTARMTVC LGKKVIHAVG PDFRKHPEAE ALKLLQNAYH AVADLVNEHN
100  IKSVAIPLLS TGIYAAGKDR LEVSLNCLTT ALDRTDADVT IYCLDKKWKE
150  RIDAALQLKE SVTELKDEDM EIDDELVWIH PDSCLKGRKG FSTTKGKLYS
200  YFEGTKFHQA AKDMAEIKVL FPNDQESNEQ LCAYILGETM EAIREKCPVD
250  HNPSSSPPKT LPCLCMYAMT PERVHRLRSN NVKEVTVCSS TPLPKHKIKN
300  VQKVQCTKVV LFNPHTPAFV PARKYI

Composition (mole %)

Ala 29 (8.9 %) Asx  0 ( 0.0 %) Cys 12 (3.7 %) Asp 18 (5.5 %) Glu 24 (7.4 %)
Phe  8 (2.5 %) Gly 13 ( 4.0 %) His 10 (3.1 %) Ile 15 (4.6 %) Lys 29 (8.9 %)
Leu 27 (8.3 %) Met  6 ( 1.8 %) Asn 14 (4.3 %) Pro 20 (6.1 %) Gln  8 (2.5 %)
Arg 16 (4.9 %) Ser 17 ( 5.2 %) Thr 23 (7.1 %) Val 24 (7.4 %) Trp  3 (0.9 %)
Tyr 10 (3.1 %) Glx  0 ( 0.0 %) xxx  0 (0.0 %)

Titration of the conserved domain (number of residues protonated)

(-) (+) net pH Cys Asp Glu Tyr COOH charge His Lys Arg NH2 charge charge
1.0 12.00 17.98 23.99 10.00 1.00 -0.03 10.00 29.00 16.00 1.00 56.00 55.97
1.5 12.00 17.93 23.96 10.00 0.99 -0.12 10.00 29.00 16.00 1.00 56.00 55.88
2.0 12.00 17.78 23.87 10.00 0.97 -0.38 10.00 29.00 16.00 1.00 56.00 55.62
2.5 12.00 17.33 23.58 10.00 0.92 -1.17 10.00 29.00 16.00 1.00 56.00 54.83
3.0 12.00 16.03 22.72 10.00 0.78 -3.47 10.00 29.00 16.00 1.00 56.00 52.53
3.5 12.00 12.96 20.38 10.00 0.53 -9.13 9.99 29.00 16.00 1.00 55.99 46.86
4.0 12.00 8.07 15.36 10.00 0.27 -19.30 9.97 29.00 16.00 1.00 55.97 36.67
4.5 12.00 3.68 8.64 10.00 0.10 -30.58 9.90 29.00 16.00 1.00 55.90 25.32
5.0 11.99 1.35 3.62 10.00 0.04 -38.00 9.69 29.00 16.00 1.00 55.69 17.69
5.5 11.98 0.45 1.28 10.00 0.01 -41.28 9.09 29.00 16.00 1.00 55.09 13.81
6.0 11.94 0.15 0.42 10.00 0.00 -42.49 7.60 29.00 16.00 1.00 53.60 11.11
6.5 11.81 0.05 0.13 10.00 0.00 -43.01 5.00 29.00 16.00 0.99 50.99 7.98
7.0 11.43 0.01 0.04 10.00 0.00 -43.52 2.40 29.00 16.00 0.97 48.37 4.85
7.5 10.36 0.00 0.01 10.00 0.00 -44.63 0.91 28.99 16.00 0.92 46.82 2.19
8.0 7.99 0.00 0.00 9.99 0.00 -47.02 0.31 28.95 16.00 0.78 46.04 -0.98
8.5 4.64 0.00 0.00 9.96 0.00 -50.40 0.10 28.85 16.00 0.53 45.48 -4.92
9.0 2.00 0.00 0.00 9.89 0.00 -53.11 0.03 28.54 15.99 0.27 44.83 -8.28
9.5 0.71 0.00 0.00 9.66 0.00 -54.63 0.01 27.59 15.98 0.10 43.68 -10.95
10.0 0.23 0.00 0.00 8.99 0.00 -55.78 0.00 24.95 15.95 0.04 40.94 -14.84
10.5 0.08 0.00 0.00 7.38 0.00 -57.54 0.00 19.17 15.84 0.01 35.02 -22.52
11.0 0.02 0.00 0.00 4.71 0.00 -60.27 0.00 11.06 15.51 0.00 26.57 -33.70
11.5 0.01 0.00 0.00 2.20 0.00 -62.79 0.00 4.73 14.55 0.00 19.28 -43.51
12.0 0.00 0.00 0.00 0.82 0.00 -64.18 0.00 1.68 12.16 0.00 13.84 -50.34
12.5 0.00 0.00 0.00 0.27 0.00 -64.73 0.00 0.55 8.00 0.00 8.55 -56.18
13.0 0.00 0.00 0.00 0.09 0.00 -64.91 0.00 0.18 3.84 0.00 4.02 -60.89

Isoelectric point

                                          (-)                                (+)    net 
 pI    Cys    Asp    Glu    Tyr   COOH   charge  His    Lys    Arg    NH2  charge  charge
7.86   8.79   0.00   0.01   9.99  0.00  -46.21   0.42  28.97  16.00  0.83  46.22    0.01

The variable domain

223 residues (5079-5747) 24660 dalton

Sequence

             10         20         30         40         50
327                              EVPE QPTAPPAQAE EAPEVVATPS
350  PSTADNTSLD VTDISLDMDD SSEGSLFSSF SGSDNSITSM DSWSSGPSSL
400  EIVDRRQVVV ADVHAVQEPA PIPPPRLKKM ARLAAARKEP TPPASNSSES
450  LHLSFGGVSM SLGSIFDGET ARQAAVQPLA TGPTDVPMSF GSFSDGEIDE
500  LSRRVTESEP VLFGSFEPGE VNSIISSRSA VSFPLRKQRR RRRSRRTEY

Composition (mole %)

Ala 19 (8.5 %) Asx  0 ( 0.0 %) Cys  0 (0.0 %) Asp 14 (6.3 %) Glu 18 (8.1 %)
Phe  9 (4.0 %) Gly 12 ( 5.4 %) His  2 (0.9 %) Ile  8 (3.6 %) Lys 4 (1.8 %)
Leu 13 (5.8 %) Met  5 ( 2.2 %) Asn  4 (1.8 %) Pro 22 (9.9 %) Gln 7 (3.1 %)
Arg 17 (7.6 %) Ser 38 (17.0 %) Thr 12 (5.4 %) Val 17 (7.6 %) Trp 1 (0.4 %)
Yal  1 (0.4 %) Glx  0 ( 0.0 %) xxx  0 (0.0 %)

Titration of the variable domain (number of residues protonated)

(-) (+) net pH Cys Asp Glu Tyr COOH charge His Lys Arg NH2 charge charge
1.0 0.00 13.98 17.99 1.00 1.00 -0.03 2.00 4.00 17.00 1.00 24.00 23.97
1.5 0.00 13.95 17.97 1.00 0.99 -0.09 2.00 4.00 17.00 1.00 24.00 23.91
2.0 0.00 13.83 17.90 1.00 0.97 -0.30 2.00 4.00 17.00 1.00 24.00 23.70
2.5 0.00 13.48 17.69 1.00 0.92 -0.91 2.00 4.00 17.00 1.00 24.00 23.09
3.0 0.00 12.47 17.04 1.00 0.78 -2.71 2.00 4.00 17.00 1.00 24.00 21.29
3.5 0.00 10.08 15.28 1.00 0.53 -7.11 2.00 4.00 17.00 1.00 24.00 16.89
4.0 0.00 6.28 11.52 1.00 0.27 -14.93 1.99 4.00 17.00 1.00 23.99 9.06
4.5 0.00 2.86 6.48 1.00 0.10 -23.56 1.98 4.00 17.00 1.00 23.98 0.42
5.0 0.00 1.05 2.72 1.00 0.04 -29.19 1.94 4.00 17.00 1.00 23.94 -5.25
5.5 0.00 0.35 0.96 1.00 0.01 -31.68 1.82 4.00 17.00 1.00 23.82 -7.86
6.0 0.00 0.11 0.31 1.00 0.00 -32.58 1.52 4.00 17.00 1.00 23.52 -9.06
6.5 0.00 0.04 0.10 1.00 0.00 -32.86 1.00 4.00 17.00 0.99 22.99 -9.87
7.0 0.00 0.01 0.03 1.00 0.00 -32.96 0.48 4.00 17.00 0.97 22.45 -10.51
7.5 0.00 0.00 0.01 1.00 0.00 -32.99 0.18 4.00 17.00 0.92 22.10 -10.89
8.0 0.00 0.00 0.00 1.00 0.00 -33.00 0.06 3.99 17.00 0.78 21.83 -11.17
8.5 0.00 0.00 0.00 1.00 0.00 -33.00 0.02 3.98 17.00 0.53 21.53 -11.47
9.0 0.00 0.00 0.00 0.99 0.00 -33.01 0.01 3.94 16.99 0.27 21.21 -11.80
9.5 0.00 0.00 0.00 0.97 0.00 -33.03 0.00 3.80 16.98 0.10 20.88 -12.15
10.0 0.00 0.00 0.00 0.90 0.00 -33.10 0.00 3.44 16.95 0.04 20.43 -12.67
10.5 0.00 0.00 0.00 0.74 0.00 -33.26 0.00 2.64 16.83 0.01 19.48 -13.78
11.0 0.00 0.00 0.00 0.47 0.00 -33.53 0.00 1.53 16.48 0.00 18.01 -15.52
11.5 0.00 0.00 0.00 0.22 0.00 -33.78 0.00 0.65 15.45 0.00 16.10 -17.68
12.0 0.00 0.00 0.00 0.08 0.00 -33.92 0.00 0.23 12.92 0.00 13.15 -20.77
12.5 0.00 0.00 0.00 0.03 0.00 -33.97 0.00 0.08 8.50 0.00 8.58 -25.39
13.0 0.00 0.00 0.00 0.01 0.00 -33.99 0.00 0.02 4.08 0.00 4.10 -29.89

Isoelectric point

                                         (-)                            (+)    net 
 pI    Cys    Asp    Glu   Tyr  COOH   charge  His   Lys   Arg   NH2  charge charge
4.53  0.00   2.72   6.22  1.00  0.10  -23.96  1.98  4.00  17.00  1.00  23.98  0.02

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Send suggestions and comments to: huang@borcim.wustl.edu WWW Department of Molecular Microbiology, Box 8230 Washington University School of Medicine St. Louis, MO 63110-1093 USA		Tel 314-362-2755 FAX 314-362-1232