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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 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 HN) (resid 3 and name HN) 3.40 1.60 0.5 ! noesy.168








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 4 and name HN) (resid 4 and name HA#) 2.80 1.00 0.5 ! noesy.4













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 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 HE) 3.40 1.60 0.5 ! noesy.162




















assign (resid 7 and name HA) (resid 7 and name HB#) 5.00 3.20 0.5 ! noesy.122











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 9 and name HN) (resid 9 and name HG#) 5.00 3.20 0.5 ! noesy.18









assign (resid 9 and name HG#) (resid 9 and name HN) 5.00 3.20 0.5 ! noesy.142





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


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 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 16


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 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


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

Figure S1

.

Figure S2

Figure S3

Figure S4

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.

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