Homework 7
Answer:
the amino acid:
MLECNELIPL FYETCPAGEN ICYEMFMVAT PKVPCERGCI DVCPESSLIV KYVCCNTDRC Q
(1) David Liu would like to know the possible functional si
Please help him to identify any biologically significant sites or patterns.
ProSite Search Result for ?
PS00005 (PDOC00005) Protein kinase C phosphorylation site.
Pattern: [ST]-x-[RK].
Matching residues 30-32: MFMVA TPK VPCER
Matching residues 57-59: YVCCN TDR CQ
PS00272 (PDOC00245) Snake toxins signature.
Pattern: G-C-x(1,3)-C-P-x(8,10)-C-C-x(2)-[PDEN].
Matching residues 38-58: VPCER GCIDVCPESSLIVKYVCCNTD RCQ
(2) Please help him to analyze the enzymatic digestion by trypsin and endoproteinase Glu-C.
(a)trypsin
1 * 10 * 20 * 30 * 40 * 50
1 MLECNELIPLFYETCPAGENICYEMFMVATPKVPCERGCIDVCPESSLIV
51 KYVCCNTDRCQ
Cleavage at residues:
Average mass
Fragment Residues Mass B&B
1 1 - 32 -/M...K/V 3701.41 -265
1 - 2 1 - 37 -/M...R/G 4286.11 -212
1 - 3 1 - 51 -/M...K/Y 5730.85 -198
1 - 4 1 - 59 -/M...R/C 6685.93 -154
1 - 5 1 - 61 -/M...Q/- 6917.21 -127
2 33 - 37 K/V...R/G 602.71 128
2 - 3 33 - 51 K/V...K/Y 2047.45 -85
2 - 4 33 - 59 K/V...R/C 3002.53 -22
2 - 5 33 - 61 K/V...Q/- 3233.81 24
3 38 - 51 R/G...K/Y 1462.75 -162
3 - 4 38 - 59 R/G...R/C 2417.84 -56
3 - 5 38 - 61 R/G...Q/- 2649.11 3
4 52 - 59 K/Y...R/C 973.10 127
4 - 5 52 - 61 K/Y...Q/- 1204.37 235
5 60 - 61 R/CQ/- 249.29 665
Sorted according to mass
Fragment Residues Mass B&B
5 60 - 61 R/CQ/- 249.29 665
2 33 - 37 K/V...R/G 602.71 128
4 52 - 59 K/Y...R/C 973.10 127
4 - 5 52 - 61 K/Y...Q/- 1204.37 235
3 38 - 51 R/G...K/Y 1462.75 -162
2 - 3 33 - 51 K/V...K/Y 2047.45 -85
3 - 4 38 - 59 R/G...R/C 2417.84 -56
3 - 5 38 - 61 R/G...Q/- 2649.11 3
2 - 4 33 - 59 K/V...R/C 3002.53 -22
2 - 5 33 - 61 K/V...Q/- 3233.81 24
1 1 - 32 -/M...K/V 3701.41 -265
1 - 2 1 - 37 -/M...R/G 4286.11 -212
1 - 3 1 - 51 -/M...K/Y 5730.85 -198
1 - 4 1 - 59 -/M...R/C 6685.93 -154
1 - 5 1 - 61 -/M...Q/- 6917.21 -127
(b)endoproteinase Glu-C
1 * 10 * 20 * 30 * 40 * 50
1 MLECNELIPLFYETCPAGENICYEMFMVATPKVPCERGCIDVCPESSLIV
51 KYVCCNTDRCQ
Cleavage at residues:
Average mass
Fragment Residues Mass B&B
1 1 - 3 -/M...E/C 391.49 -600
1 - 2 1 - 6 -/M...E/L 737.85 -6
1 - 3 1 - 13 -/M...E/T 1613.92 -569
1 - 4 1 - 19 -/M...E/N 2172.53 -262
1 - 5 1 - 24 -/M...E/M 2795.23 -254
1 - 6 1 - 36 -/M...E/R 4129.92 -237
1 - 7 1 - 45 -/M...E/S 5103.06 -168
1 - 8 1 - 61 -/M...Q/- 6917.21 -127
2 4 - 6 E/C...E/L 364.38 586
2 - 3 4 - 13 E/C...E/T 1240.44 -560
2 - 4 4 - 19 E/C...E/N 1799.06 -199
2 - 5 4 - 24 E/C...E/M 2421.76 -205
2 - 6 4 - 36 E/C...E/R 3756.45 -204
2 - 7 4 - 45 E/C...E/S 4729.59 -137
2 - 8 4 - 61 E/C...Q/- 6543.73 -103
3 7 - 13 E/L...E/T 894.08 -1051
3 - 4 7 - 19 E/L...E/N 1452.69 -380
3 - 5 7 - 24 E/L...E/M 2075.39 -337
3 - 6 7 - 36 E/L...E/R 3410.08 -284
3 - 7 7 - 45 E/L...E/S 4383.22 -193
3 - 8 7 - 61 E/L...Q/- 6197.37 -140
4 14 - 19 E/T...E/N 576.63 401
4 - 5 14 - 24 E/T...E/M 1199.33 117
4 - 6 14 - 36 E/T...E/R 2534.02 -50
4 - 7 14 - 45 E/T...E/S 3507.16 -5
4 - 8 14 - 61 E/T...Q/- 5321.30 -8
5 20 - 24 E/N...E/M 640.72 -224
5 - 6 20 - 36 E/N...E/R 1975.41 -210
5 - 7 20 - 45 E/N...E/S 2948.55 -100
5 - 8 20 - 61 E/N...Q/- 4762.69 -66
6 25 - 36 E/M...E/R 1352.71 -204
6 - 7 25 - 45 E/M...E/S 2325.85 -70
6 - 8 25 - 61 E/M...Q/- 4139.99 -45
7 37 - 45 E/R...E/S 991.16 107
7 - 8 37 - 61 E/R...Q/- 2805.30 30
8 46 - 61 E/S...Q/- 1832.16 -12
Sorted according to mass
Fragment Residues Mass B&B
2 4 - 6 E/C...E/L 364.38 586
1 1 - 3 -/M...E/C 391.49 -600
4 14 - 19 E/T...E/N 576.63 401
5 20 - 24 E/N...E/M 640.72 -224
1 - 2 1 - 6 -/M...E/L 737.85 -6
3 7 - 13 E/L...E/T 894.08 -1051
7 37 - 45 E/R...E/S 991.16 107
4 - 5 14 - 24 E/T...E/M 1199.33 117
2 - 3 4 - 13 E/C...E/T 1240.44 -560
6 25 - 36 E/M...E/R 1352.71 -204
3 - 4 7 - 19 E/L...E/N 1452.69 -380
1 - 3 1 - 13 -/M...E/T 1613.92 -569
2 - 4 4 - 19 E/C...E/N 1799.06 -199
8 46 - 61 E/S...Q/- 1832.16 -12
5 - 6 20 - 36 E/N...E/R 1975.41 -210
3 - 5 7 - 24 E/L...E/M 2075.39 -337
1 - 4 1 - 19 -/M...E/N 2172.53 -262
6 - 7 25 - 45 E/M...E/S 2325.85 -70
2 - 5 4 - 24 E/C...E/M 2421.76 -205
4 - 6 14 - 36 E/T...E/R 2534.02 -50
1 - 5 1 - 24 -/M...E/M 2795.23 -254
7 - 8 37 - 61 E/R...Q/- 2805.30 30
5 - 7 20 - 45 E/N...E/S 2948.55 -100
3 - 6 7 - 36 E/L...E/R 3410.08 -284
4 - 7 14 - 45 E/T...E/S 3507.16 -5
2 - 6 4 - 36 E/C...E/R 3756.45 -204
1 - 6 1 - 36 -/M...E/R 4129.92 -237
6 - 8 25 - 61 E/M...Q/- 4139.99 -45
3 - 7 7 - 45 E/L...E/S 4383.22 -193
2 - 7 4 - 45 E/C...E/S 4729.59 -137
5 - 8 20 - 61 E/N...Q/- 4762.69 -66
1 - 7 1 - 45 -/M...E/S 5103.06 -168
4 - 8 14 - 61 E/T...Q/- 5321.30 -8
3 - 8 7 - 61 E/L...Q/- 6197.37 -140
2 - 8 4 - 61 E/C...Q/- 6543.73 -103
1 - 8 1 - 61 -/M...Q/- 6917.21 -127
(3) Calculate the molecular weight of this protein.
Average mass: M=6917.21
Monoisotopic mass: M=6912.08
