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Supplementary Material
Table S1. 1H NMR chemical shifts (ppm) for 1 mM of Cn-AMP1 in 100 mM of SDS-
d25 at pH 3.97 and 25 °C.
Residues HN Hα Hβ Others
Ser-1 - 4.33 4.27 4.06(γH)
Val-2 8.64 3.95 2.17 1.03(γCH3). 0.96(γCH3)
Ala-3 8.23 4.16 1.39 -
Gly-4 8.09 3.89 - -
Arg-5 7.95 4.22 1.72 1.91(βH1). 1.71(γCH2). 3.18(δCH2). 7.16(εNH)
Ala-6 8.15 4.24 1.43 -
Gln-7 7.92 4.25 2.03 2.15(βH1). 2.41(γH)
Gly-8 8.14 3.96 - -
Met-9 7.79 4.27 1.99 2.09 (βH1). 2.54 (γH)
Table S2. 1H NMR chemical shifts (ppm) for 1 mM of Cn-AMP1 without SDS-d25 at
pH 3.97 and 25°C.
Residues HN Hα Hβ Others
Ser-1 - - 3.88 -
Val-2 8.56 4.13 2.00 0.92 (γCH3), 0.87 (γCH3)
Ala-3 8.48 4.25 1.33 -
Gly-4 8.35 3.88 - -
Arg-5 8.11 4.29 1.73 1.68 (βH1), 1.56 (γCH2), 3.13 (δCH2), 7.16
(εNH)
Ala-6 8.40 4.25 1.33 -
Gln-7 8.36 4.22 2.00 1.96 (βH1), 2.33 (γH)
Gly-8 8.44 3.88 - -
Met-9 7.86 4.31 2.07 1.92 (βH1), 2.45 (γH)
Table S3. Signal area for i-i+3 and NH-NH NOE interactions.
Type NOE Integral
i – i+3 4HN – 1HA 1598.01
i – i+3 5HN – 2HA 93.69
i – i+3 6HN – 3HA 919.22
i – i+3 8HN – 5HA 2288.87
NH – NH 3HN – 2HN 931.62
NH – NH 4HN – 3HN 1198.99
NH – NH 5HN – 4HN 2341.77
NH – NH 6HN – 5HN 1960.97
NH – NH 7HN – 6HN 3238.27
NH – NH 8HN – 7HN 1960.97
NH – NH 9HN – 8HN 610.33
Table S4. Constraints data of Cn-AMP1 peptide in SDS-d25 micelles, pH 3.97 and 25
°C.
assign (resid 1 and name HT#) (resid 1 and name HA) 5.00 3.20 0.5! noesy.7
assign (resid 1 and name HB#) (resid 3 and name HB#) 5.00 3.20 0.5! noesy.75
assign (resid 1 and name HB#) (resid 1 and name HG) 3.40 1.60 0.5 ! noesy.81
assign (resid 1 and name HB#) (resid 2 and name HB) 5.00 3.20 0.5 ! noesy.121
assign (resid 2 and name HN) (resid 2 and name HA) 3.40 1.60 0.5 ! noesy.0
assign (resid 2 and name HN) (resid 2 and name HB) 2.80 1.00 0.5 ! noesy.8
assign (resid 2 and name HN) (resid 2 and name HG##) 3.40 1.60 0.5 ! noesy.29
assign (resid 2 and name HN) (resid 1 and name HA) 3.40 1.60 0.5 ! noesy.32
assign (resid 2 and name HN) (resid 1 and name HB#) 3.40 1.60 0.5 ! noesy.33
assign (resid 2 and name HN) (resid 1 and name HG) 5.00 3.20 0.5 ! noesy.35
assign (resid 2 and name HA) (resid 2 and name HG##) 3.40 1.60 0.5 ! noesy.77
assign (resid 2 and name HA) (resid 2 and name HB) 5.00 3.20 0.5 ! noesy.80
assign (resid 2 and name HB) (resid 1 and name HB#) 5.00 3.20 0.5 ! noesy.97
assign (resid 2 and name HB) (resid 2 and name HA) 5.00 3.20 0.5 ! noesy.98
assign (resid 2 and name HG##) (resid 2 and name HA) 3.40 1.60 0.5 ! noesy.109
assign (resid 2 and name HA) (resid 5 and name HN) 3.40 1.60 0.5 ! noesy.129
assign (resid 2 and name HB) (resid 2 and name HN) 2.80 1.00 0.5 ! noesy.138
assign (resid 2 and name HB) (resid 3 and name HN) 3.40 1.60 0.5 ! noesy.139
assign (resid 2 and name HG##) (resid 2 and name HN) 3.40 1.60 0.5 ! noesy.156
assign (resid 2 and name HG##) (resid 3 and name HN) 5.00 3.20 0.5 ! noesy.158
assign (resid 2 and name HG##) (resid 4 and name HN) 5.00 3.20 0.5 ! noesy.160
assign (resid 2 and name HA) (resid 2 and name HN) 3.40 1.60 0.5 ! noesy.163
assign (resid 2 and name HN) (resid 3 and name HN) 3.40 1.60 0.5 ! noesy.168
assign (resid 3 and name HN) (resid 3 and name HA) 2.80 1.00 0.5 ! noesy.1
assign (resid 3 and name HN) (resid 3 and name HB#) 2.80 1.00 0.5 ! noesy.10
assign (resid 3 and name HN) (resid 1 and name HA) 5.00 3.20 0.5 ! noesy.38
assign (resid 3 and name HN) (resid 1 and name HB#) 5.00 3.20 0.5 ! noesy.39
assign (resid 3 and name HN) (resid 2 and name HA) 3.40 1.60 0.5 ! noesy.41
assign (resid 3 and name HN) (resid 2 and name HB) 3.40 1.60 0.5 ! noesy.42
assign (resid 3 and name HN) (resid 2 and name HG##) 5.00 3.20 0.5 ! noesy.43
assign (resid 3 and name HB#) (resid 1 and name HB#) 5.00 3.20 0.5 ! noesy.107
assign (resid 3 and name HB#) (resid 3 and name HN) 2.80 1.00 0.5 ! noesy.151
assign (resid 3 and name HB#) (resid 5 and name HN) 5.00 3.20 0.5 ! noesy.155
assign (resid 3 and name HA) (resid 3 and name HN) 2.80 1.00 0.5 ! noesy.164
assign (resid 3 and name HN) (resid 2 and name HN) 3.40 1.60 0.5 ! noesy.170
assign (resid 3 and name HN) (resid 4 and name HN) 3.40 1.60 0.5 ! noesy.171
assign (resid 3 and name HN) (resid 1 and name HG) 5.00 3.20 0.5 ! noesy.189
assign (resid 4 and name HN) (resid 4 and name HA#) 2.80 1.00 0.5 ! noesy.4
assign (resid 4 and name HN) (resid 3 and name HA) 3.40 1.60 0.5 ! noesy.45
assign (resid 4 and name HN) (resid 1 and name HB#) 5.00 3.20 0.5 ! noesy.56
assign (resid 4 and name HN) (resid 2 and name HB) 5.00 3.20 0.5 ! noesy.57
assign (resid 4 and name HN) (resid 6 and name HB#) 5.00 3.20 0.5 ! noesy.60
assign (resid 4 and name HN) (resid 2 and name HG##) 5.00 3.20 0.5 ! noesy.61
assign (resid 4 and name HN) (resid 5 and name HN) 2.80 1.00 0.5 ! noesy.174
assign (resid 4 and name HN) (resid 3 and name HN) 3.40 1.60 0.5 ! noesy.175
assign (resid 4 and name HN) (resid 1 and name HA) 5.00 3.20 0.5 ! noesy.190
assign (resid 4 and name HN) (resid 1 and name HG) 5.00 3.20 0.5 ! noesy.195
assign (resid 4 and name HN) (resid 2 and name HA) 3.40 1.60 0.5 ! noesy.196
assign (resid 5 and name HN) (resid 5 and name HA) 3.40 1.60 0.5 ! noesy.5
assign (resid 5 and name HN) (resid 5 and name HB#) 5.00 3.20 0.5 ! noesy.13
assign (resid 5 and name HN) (resid 5 and name HG#) 5.00 3.20 0.5 ! noesy.14
assign (resid 5 and name HE) (resid 5 and name HA) 5.00 3.20 0.5 ! noesy.21
assign (resid 5 and name HE) (resid 5 and name HG#) 2.80 1.00 0.5 ! noesy.22
assign (resid 5 and name HE) (resid 5 and name HB#) 3.40 1.60 0.5 ! noesy.23
assign (resid 5 and name HN) (resid 2 and name HA) 3.40 1.60 0.5 ! noesy.62
assign (resid 5 and name HN) (resid 6 and name HB#) 5.00 3.20 0.5 ! noesy.68
assign (resid 5 and name HN) (resid 3 and name HB#) 5.00 3.20 0.5 ! noesy.69
assign (resid 5 and name HG#) (resid 5 and name HB#) 2.80 1.00 0.5 ! noesy.83
assign (resid 5 and name HB#) (resid 8 and name HA#) 5.00 3.20 0.5 ! noesy.100
assign (resid 5 and name HB#) (resid 5 and name HG#) 2.80 1.00 0.5 ! noesy.101
assign (resid 5 and name HG#) (resid 8 and name HA#) 5.00 3.20 0.5 ! noesy.119
assign (resid 5 and name HG#) (resid 5 and name HE) 2.80 1.00 0.5 ! noesy.137
assign (resid 5 and name HB#) (resid 5 and name HN) 5.00 3.20 0.5 ! noesy.149
assign (resid 5 and name HB#) (resid 8 and name HN) 5.00 3.20 0.5 ! noesy.150
assign (resid 5 and name HB#) (resid 5 and name HE) 3.40 1.60 0.5 ! noesy.162
assign (resid 5 and name HN) (resid 4 and name HN) 2.80 1.00 0.5 ! noesy.180
assign (resid 5 and name HN) (resid 3 and name HA) 5.00 3.20 0.5 ! noesy.197
assign (resid 5 and name HN) (resid 1 and name HA) 5.00 3.20 0.5 ! noesy.198
assign (resid 5 and name HN) (resid 4 and name HA#) 5.00 3.20 0.5 ! noesy.202
assign (resid 6 and name HN) (resid 6 and name HA) 2.80 1.00 0.5 ! noesy.2
assign (resid 6 and name HN) (resid 2 and name HA) 5.00 3.20 0.5 ! noesy.47
assign (resid 6 and name HN) (resid 5 and name HB#) 5.00 3.20 0.5 ! noesy.51
assign (resid 6 and name HA) (resid 7 and name HN) 5.00 3.20 0.5 ! noesy.124
assign (resid 6 and name HB#) (resid 8 and name HN) 5.00 3.20 0.5 ! noesy.152
assign (resid 6 and name HB#) (resid 4 and name HN) 5.00 3.20 0.5 ! noesy.153
assign (resid 6 and name HB#) (resid 5 and name HN) 5.00 3.20 0.5 ! noesy.154
assign (resid 6 and name HN) (resid 3 and name HN) 5.00 3.20 0.5 ! noesy.176
assign (resid 6 and name HN) (resid 3 and name HA) 5.00 3.20 0.5 ! noesy.191
assign (resid 6 and name HN) (resid 4 and name HA#) 5.00 3.20 0.5 ! noesy.208
assign (resid 6 and name HN) (resid 5 and name HA) 3.40 1.60 0.5 ! noesy.213
assign (resid 6 and name HN) (resid 5 and name HN) 2.80 1.00 0.5 ! noesy.214
assign (resid 7 and name HN) (resid 7 and name HA) 2.80 1.00 0.5 ! noesy.6
assign (resid 7 and name HN) (resid 7 and name HB#) 5.00 3.20 0.5 ! noesy.17
assign (resid 7 and name HG#) (resid 7 and name HB#) 5.00 3.20 0.5 ! noesy.89
assign (resid 7 and name HA) (resid 7 and name HB#) 5.00 3.20 0.5 ! noesy.122
assign (resid 7 and name HA) (resid 7 and name HN) 2.80 1.00 0.5 ! noesy.125
assign (resid 7 and name HN) (resid 6 and name HN) 3.40 1.60 0.5 ! noesy.181
assign (resid 7 and name HN) (resid 9 and name HN) 5.00 3.20 0.5 ! noesy.187
assign (resid 7 and name HN) (resid 6 and name HA) 5.00 3.20 0.5 ! noesy.203
assign (resid 7 and name HN) (resid 5 and name HA) 5.00 3.20 0.5 ! noesy.209
assign (resid 8 and name HN) (resid 8 and name HA#) 2.80 1.00 0.5 ! noesy.3
assign (resid 8 and name HN) (resid 6 and name HB#) 5.00 3.20 0.5 ! noesy.11
assign (resid 8 and name HN) (resid 7 and name HA) 5.00 3.20 0.5 ! noesy.46
assign (resid 8 and name HN) (resid 7 and name HB#) 5.00 3.20 0.5 ! noesy.49
assign (resid 8 and name HN) (resid 5 and name HB#) 5.00 3.20 0.5 ! noesy.52
assign (resid 8 and name HA#) (resid 9 and name HN) 5.00 3.20 0.5 ! noesy.126
assign (resid 8 and name HA#) (resid 8 and name HN) 2.80 1.00 0.5 ! noesy.133
assign (resid 8 and name HN) (resid 7 and name HN) 2.80 1.00 0.5 ! noesy.172
assign (resid 8 and name HN) (resid 9 and name HN) 5.00 3.20 0.5 ! noesy.173
assign (resid 8 and name HN) (resid 5 and name HA) 5.00 3.20 0.5 ! noesy.200
assign (resid 8 and name HN) (resid 6 and name HA) 5.00 3.20 0.5 ! noesy.205
assign (resid 9 and name HN) (resid 9 and name HA) 5.00 3.20 0.5 ! noesy.25
assign (resid 9 and name HN) (resid 9 and name HG#) 5.00 3.20 0.5 ! noesy.18
assign (resid 9 and name HN) (resid 9 and name HB#) 5.00 3.20 0.5 ! noesy.19
assign (resid 9 and name HN) (resid 7 and name HB#) 5.00 3.20 0.5 ! noesy.20
assign (resid 9 and name HN) (resid 8 and name HA#) 5.00 3.20 0.5 ! noesy.66
assign (resid 9 and name HN) (resid 5 and name HB#) 5.00 3.20 0.5 ! noesy.70
assign (resid 9 and name HG#) (resid 9 and name HB#) 5.00 3.20 0.5 ! noesy.86
assign (resid 9 and name HG#) (resid 7 and name HB#) 5.00 3.20 0.5 ! noesy.88
assign (resid 9 and name HB#) (resid 7 and name HG#) 5.00 3.20 0.5 ! noesy.102
assign (resid 9 and name HB#) (resid 9 and name HG#) 5.00 3.20 0.5 ! noesy.113
assign (resid 9 and name HG#) (resid 9 and name HN) 5.00 3.20 0.5 ! noesy.142
assign (resid 9 and name HB#) (resid 9 and name HN) 5.00 3.20 0.5 ! noesy.145
assign (resid 9 and name HN) (resid 8 and name HN) 5.00 3.20 0.5 ! noesy.179
assign (resid 9 and name HN) (resid 7 and name HA) 5.00 3.20 0.5 ! noesy.194
assign (resid 9 and name HN) (resid 5 and name HA) 5.00 3.20 0.5 ! noesy.206
Table S5. H-bonds occurrence and geometric distances in the final 20 lowest-energy
structures of 1mM Cn-AMP1 peptide in presence of 100 mM SDS-d25 micelles, at
25°C and pH 3.97.
Peptide Donor Acceptor ParametersDd-a Dh-a A(d-H-N) A(a-O=C)
All
R5 (N) S1 (O) 3.14 2.41 132.05 153.36R5 (N) V2 (O) 2.90 2.09 140.18 124.03A6 (N) V2 (O) 2.81 2.15 123.86 168.58M9 (N) G4 (O) 3.20 2.27 160.12 165.99
Model 1
G4 (N) S1 (O) 3.08 2.12 166.60 129.08R5 (N) V2 (O) 2.82 1.97 145.32 132.17A6 (N) V2 (O) 3.20 2.58 121.38 157.81M9 (N) G4 (O) 3.35 2.41 161.61 157.54
Model 2
G4 (N) S1 (O) 3.41 2.57 144.86 104.88R5 (N) V2 (O) 3.07 2.26 139.47 123.50A6 (N) V2 (O) 2.64 1.99 122.40 163.08M9 (N) G4 (O) 2.98 2.09 152.38 164.56
Model 3
G4 (N) S1 (O) 3.09 2.16 160.27 119.53R5 (N) V2 (O) 2.93 2.08 145.77 125.84A6 (N) V2 (O) 3.13 2.49 123.36 161.49M9 (N) G4 (O) 3.07 2.18 152.71 159.00A3 (N) S1 (OG) 3.26 2.46 139.23 999.99*
Model 4
G4 (N) S1 (O) 2.90 2.06 143.32 116.85R5 (N) V2 (O) 3.17 2.36 140.80 119.74A6 (N) V2 (O) 3.38 2.89 112.71 149.14A6 (N) A3 (O) 3.06 2.18 149.36 102.37M9 (N) G4 (O) 3.08 2.24 145.16 155.23Q7 (NE2) A3 (O) 2.61 1.99 114.93 151.54Q7 (NE2) A3 (O) 2.61 2.61 78.26 151.54Q7 (NE2) G4 (O) 2.81 3.30 53.69 103.00Q7 (NE2) G4 (O) 2.81 1.79 160.38 103.00Q7 (NE2) M9 (O) 3.20 3.77 49.90 159.37Q7 (NE2) M9 (O) 3.20 2.77 104.41 159.37
Model 5
G4 (N) S1 (O) 2.93 2.08 144.11 112.39R5 (N) S1 (O) 3.15 2.37 136.37 162.50R5 (N) V2 (O) 3.39 2.61 137.81 105.68A6 (N) V2 (O) 3.00 2.42 118.32 153.77M9 (N) G4 (O) 3.25 2.29 168.00 166.58G4 (N) S1 (OG) 3.36 2.78 119.32 999.99*
Model 6R5 (N) V2 (O) 2.90 2.08 142.29 126.30A6 (N) V2 (O) 3.06 2.49 117.46 160.99
M9 (N) G4 (O) 3.05 2.21 143.32 162.79Q7 (NE2) M9 (O) 3.37 3.98 47.99 135.57Q7 (NE2) M9 (O) 3.37 2.73 118.75 135.57
Model 7R5 (N) V2 (O) 2.85 2.02 142.53 129.48A6 (N) V2 (O) 3.09 2.55 115.27 159.15M9 (N) G4 (O) 2.96 2.09 148.20 165.87
Model 8G4 (N) S1 (O) 3.17 2.32 145.66 107.65R5 (N) V2 (O) 3.01 2.12 151.10 131.30A6 (N) A3 (O) 3.01 2.17 144.94 103.48
Model 9R5 (N) V2 (O) 3.03 2.13 153.14 130.30A6 (N) V2 (O) 3.43 2.84 120.17 149.25M9 (N) G4 (O) 3.37 2.41 168.12 162.52
Model 10
R5 (N) S1 (O) 3.14 2.41 132.05 153.36R5 (N) V2 (O) 2.90 2.09 140.18 124.03A6 (N) V2 (O) 2.81 2.15 123.86 168.58M9 (N) G4 (O) 3.20 2.27 160.12 165.99
Model 11
G4 (N) S1 (O) 2.98 2.13 144.38 112.15R5 (N) V2 (O) 3.34 2.52 143.33 113.10A6 (N) V2 (O) 3.22 2.67 116.36 151.91M9 (N) G4 (O) 2.98 2.07 154.74 166.24Q7 (NE2) A3 (O) 2.64 2.29 97.11 160.85Q7 (NE2) A3 (O) 2.64 2.48 86.54 160.85
Model 12
G4 (N) S1 (O) 2.91 2.05 147.48 118.33R5 (N) V2 (O) 3.24 2.40 145.12 120.71A6 (N) A3 (O) 2.98 2.11 148.22 104.68M9 (N) G4 (O) 3.13 2.23 153.89 153.15
Model 13
G4 (N) S1 (O) 3.08 2.32 134.48 105.99R5 (N) S1 (O) 3.07 2.33 132.34 156.24R5 (N) V2 (O) 2.85 2.07 136.64 123.22A6 (N) V2 (O) 2.92 2.24 125.89 169.40M9 (N) G4 (O) 3.15 2.25 154.25 161.23
Model 14
G4 (N) S1 (O) 2.93 2.05 150.09 121.62R5 (N) V2 (O) 2.94 2.21 131.01 120.22A6 (N) V2 (O) 2.75 2.13 121.05 169.07M9 (N) G4 (O) 3.27 2.33 162.31 161.85Q7 (NE2) A3 (O) 2.59 2.26 95.83 153.33Q7 (NE2) A3 (O) 2.59 2.37 89.48 153.33Q7 (NE2) G4 (O) 3.23 3.86 46.87 98.10Q7 (NE2) G4 (O) 3.23 2.20 163.26 98.10
Model 15R5 (N) V2 (O) 3.01 2.13 149.34 128.58A6 (N) V2 (O) 3.22 2.62 120.29 153.49M9 (N) G4 (O) 2.86 1.97 151.51 166.84
Model 16
G4 (N) S1 (O) 3.01 2.16 144.80 111.29R5 (N) V2 (O) 3.06 2.16 154.08 131.18A6 (N) V2 (O) 3.08 2.35 131.82 153.22M9 (N) G4 (O) 3.11 2.20 155.13 164.48
Model 17
G4 (N) S1 (O) 2.89 2.03 146.63 114.60R5 (N) S1 (O) 3.48 2.71 136.51 159.83R5 (N) V2 (O) 3.44 2.62 142.84 108.56A6 (N) V2 (O) 3.13 2.53 120.79 151.76M9 (N) G4 (O) 3.22 2.29 161.59 165.20
Model 18R5 (N) V2 (O) 2.98 2.10 150.68 129.32A6 (N) V2 (O) 3.28 2.67 121.05 154.46M9 (N) G4 (O) 3.12 2.19 159.61 166.25
Model 19
G4 (N) S1 (O) 3.08 2.31 135.56 106.03R5 (N) S1 (O) 3.35 2.62 132.43 153.24R5 (N) V2 (O) 2.89 2.05 143.26 127.16A6 (N) V2 (O) 2.98 2.36 121.51 162.40M9 (N) G4 (O) 3.23 2.28 164.55 163.76G4 (N) S1 (OG) 3.43 2.86 118.85 999.99*
Model 20
G4 (N) S1 (O) 3.05 2.24 139.51 107.82R5 (N) V2 (O) 3.03 2.14 152.76 131.05A6 (N) V2 (O) 3.03 2.27 134.60 154.16M9 (N) G4 (O) 3.05 2.15 155.18 165.77
Dd-a = Distance Between Donor and AcceptorDh-a = Distance Between Hydrogen and AcceptorA(d-H-N) = Angle Between Donor-H-N A(a-O=C) = Angle Between Acceptor-O=C*Angle undefined
Supplementary Figure Legends
Figure S1. TOCSY experiments for 1 mM of Cn-AMP1 peptide at 25 °C and pH 3.97
(A) in 100 mmol.L-1 of SDS-d25 and (B) without SDS-d25.
Figure S2. Hα-HN NOESY for 1 mM of Cn-AMP1 peptide at 25 °C and pH 3.97 (A) in
100 mmol.L-1 of SDS-d25 and (B) without SDS-d25. NOE chemical shift assignments
were carefully performed for the peptide using standard sequential assignment protocols
(Wüthrich, K. NMR of proteins and nucleic acids, 1986, Wiley, New York) and
NMRview software (Johnson B., Blevins RA, J. Biomol. NMR, 1994, 4, 603-614).
Highlighted correlations refer to interactions of type i, i +2, i +3 and i, i +4, suggesting a
helical conformation of the peptide in the presence of SDS-d25.
Figure S3. Distribution of the main inter-residue NOEs observed for each residue of
Cn-AMP1 peptide. The correlations were obtained from the NOESY spectrum of 1 mM
of Cn-AMP1 peptide at 25 °C and pH 3.97 and organized according to the atoms
involved.
Figure S4. Ramachandran diagram for the twenty lower energy structures for Cn-
AMP1 peptide at 1 mM and 25 °C, with 100 mM of SDS-d25 and pH 3.97. Regions: A)
favorable; B) allowed; C) generously allowed and D) forbidden. In C region, the Gly
residues are highlighted.