Structure-activity Relationship of Antimicrobial Peptide SAMP1 and Its Analog Peptides†

doi:10.7503/cjcu20180680

Abstract

Abstract:

The synthetic antimicrobial peptide 1(SAMP1) was used as the research template, eight SAMP1 analog peptides were designed and synthesized by amino acid sequence rearrangement, different positive charged amino acids replacement and hydrophobic amino acid substitutions. Bioinformatics software was used to predict the physicochemical properties of SAMP1 and its analog peptides. Secondary structures were determined by circular dichroism(CD) technique. Antimicrobial activity was determined by 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazoliumbromide(MTT) assay. Hemolytic properties of peptides were evaluated by erythrocyte hemolysis assay. The results showed that most of the analogues of SAMP1 had low hemolytic toxicity, and high broad-spectrum antimicrobial activity. The results of CD showed that their secondary structures were mainly α-helix and random coil, the proportion of α-helix increased in the 50% volume fraction of 2,2,2-trifluoroethanol(TFE) solution. Compared with mother peptide SAMP1, the antimicrobial activities of the analog peptides SAMP1-A1, SAMP1-A2 and SAMP1-A3, which were obtained after sequence rearrangement, did not change so much. While the hemolytic toxicity of SAMP1-A2, in which uniform distribution of positive charged amino acids in the sequence, increased. The antimicrobial activity of the peptide SAMP1-A4, which was obtained by substituting arginine(Arg) for lysine(Lys) in the SAMP1 sequence, improved with less hemolytic toxicity. The antimicrobial activity of the peptides SAMP1-A5 and SAMP1-A7, obtained by replacing the hydrophobic amino acids in SAMP1 with strong hydrophobic amino acids isoleucine(Ile) and valine(Val), drastically reduced. Both of the antibacterial activity and the hemolytic toxicity of the peptide SAMP1-A8 increased, in which less strong hydrophobic amino acid tryptophan(Trp) replaced the hydrophobic amino acids in SAMP1.

Key words: Antimicrobial peptide, Structure-activity relationship, Antimicrobial activity, Hemolytic toxicity

CLC Number:

O629

TrendMD:

CHANG Junpeng,ZHAO Jiarui,CHEN Sijia,MENG Kai,SHI Weini,LI Ruifang. Structure-activity Relationship of Antimicrobial Peptide SAMP1 and Its Analog Peptides^†[J]. Chem. J. Chinese Universities, 2019, 40(4): 705.

Figures/Tables 6

References 23

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Peptide	Sequence^*	Molecular mass		t_R/min
Peptide	Sequence^*	Calculated	Observed	t_R/min
SAMP1	VRLLKKKI	1218.57	1218.2	10.5
SAMP1-A1	VLLIRKKK	1218.57	1218.4	10.5
SAMP1-A2	RVKLKLKI	1218.57	1218.4	10.5
SAMP1-A3	RKKKVLLI	1218.57	1218.4	10.6
SAMP1-A4	VRLLRRRI	1302.64	1302.2	10.3
SAMP1-A5	IRIIKKKI	1233.60	1232.2	12.7
SAMP1-A6	LRLLKKKL	1233.60	1232.2	9.8
SAMP1-A7	VRVVKKKV	1176.49	1175.2	11.4
SAMP1-A8	WRWWKKKW	1524.83	1524.2	8.4

Peptide	Sequence^*	Molecular mass		t_R/min
Peptide	Sequence^*	Calculated	Observed	t_R/min
SAMP1	VRLLKKKI	1218.57	1218.2	10.5
SAMP1-A1	VLLIRKKK	1218.57	1218.4	10.5
SAMP1-A2	RVKLKLKI	1218.57	1218.4	10.5
SAMP1-A3	RKKKVLLI	1218.57	1218.4	10.6
SAMP1-A4	VRLLRRRI	1302.64	1302.2	10.3
SAMP1-A5	IRIIKKKI	1233.60	1232.2	12.7
SAMP1-A6	LRLLKKKL	1233.60	1232.2	9.8
SAMP1-A7	VRVVKKKV	1176.49	1175.2	11.4
SAMP1-A8	WRWWKKKW	1524.83	1524.2	8.4

Peptide	pI	GRAVY	Instability index	Fat coefficient	Half-life/h
Peptide	pI	GRAVY	Instability index	Fat coefficient	Lactating cell	Yeast cell	E. coli
SAMP1	11.26	0.013	-12.48	182.5	100	>20	>10
SAMP1-A1	11.26	0.013	8.75	182.5	100	>20	>10
SAMP1-A2	11.26	0.013	-47.91	182.5	1	0.03	0.03
SAMP1-A3	11.26	0.013	-1.86	182.5	1	0.03	0.03
SAMP1-A4	12.48	-0.212	176.02	182.5	100	>20	>10
SAMP1-A5	11.26	0.225	-12.48	195	20	0.5	>10
SAMP1-A6	11.26	-0.125	11.60	195	5.5	0.05	0.05
SAMP1-A7	11.26	0.075	-5.46	145	100	>20	>10
SAMP1-A8	11.26	-2.475	80.35	0	2.8	0.05	0.05

Peptide	pI	GRAVY	Instability index	Fat coefficient	Half-life/h
Peptide	pI	GRAVY	Instability index	Fat coefficient	Lactating cell	Yeast cell	E. coli
SAMP1	11.26	0.013	-12.48	182.5	100	>20	>10
SAMP1-A1	11.26	0.013	8.75	182.5	100	>20	>10
SAMP1-A2	11.26	0.013	-47.91	182.5	1	0.03	0.03
SAMP1-A3	11.26	0.013	-1.86	182.5	1	0.03	0.03
SAMP1-A4	12.48	-0.212	176.02	182.5	100	>20	>10
SAMP1-A5	11.26	0.225	-12.48	195	20	0.5	>10
SAMP1-A6	11.26	-0.125	11.60	195	5.5	0.05	0.05
SAMP1-A7	11.26	0.075	-5.46	145	100	>20	>10
SAMP1-A8	11.26	-2.475	80.35	0	2.8	0.05	0.05

Peptide	Solution	α-Helix(%)	β-Sheet(%)	β-Turn(%)	Random(%)
SAMP1	PBS	54.8	6.1	16.9	22.1
	SDS	39.0	6.9	20.5	33.7
	TFE	56.3	5.1	16.6	22.0
SAMP1-A1	PBS	20.6	24.7	19.0	35.7
	SDS	54.7	5.8	17.0	22.3
	TFE	55.0	5.7	16.9	22.3
SAMP1-A2	PBS	34.2	13.5	22.2	30.0
	SDS	18.6	29.4	22.0	30.0
	TFE	55.0	5.7	16.9	22.3
SAMP1-A3	PBS	34.2	13.5	22.2	30.0
	SDS	18.6	29.4	22.0	30.0
	TFE	66.0	2.1	10.6	21.1
SAMP1-A4	PBS	19.7	23.9	20.0	36.3
	SDS	42.4	6.7	29.2	21.6
	TFE	80.8	0.5	5.0	14.3
SAMP1-A5	PBS	31.9	15.5	21.7	31.0
	SDS	74.0	3.4	6.0	16.5
	TFE	79.8	0.4	5.4	14.5
SAMP1-A6	PBS	33.6	7.4	21.9	37.1
	SDS	78.8	0.7	5.2	15.2
	TFE	80.7	0.9	5.0	14.4
SAMP1-A7	PBS	55.1	5.5	17.1	22.3
	SDS	56.2	5.1	16.6	22.0
	TFE	56.4	5.0	16.5	22.0
SAMP1-A8	PBS	20.9	26.1	21.7	31.3
	SDS	25.9	16.9	17.6	39.7
	TFE	9.3	45.9	10.8	34.1