Design and Bioactivity Evaluation of Long-effective Modified Melittin (GPG) with Antitumor and Antibacterial Function

doi:10.11924/j.issn.1000-6850.casb20200200116

Abstract

Abstract:

The purpose is to improve the clinical application value of melittin. A series of polypeptide sequences were designed by hybridpeptide method. Based on the structure-activity relationship,we used the molecular structure of amphiphilic α-helix cationic peptide as the design template, to get the long-effective modified melittin with low toxicity, antitumor and antibacterial activity. Their bioactivities were compared and screened first by bioinformatics tools, then chemical synthesis was performed, and the bioactivities of the synthetic peptides were compared to obtain the designed target peptide (GPG). The experiment showed that GPG had long-term efficacy. When the concentration of GPG was 150 μmol/L, its hemolytic ratio was still zero, when the concentration was 33.3 μmol/L, the tumor inhibition ratio of H₂₂, SMMC-7721 and SW-1116 tumor cells was (78.0±0.6)%, (74.0±1.2)% and (74.0±0.5)%, respectively. When the concentration was 100 μmol/L, the diameter of inhibition zone on Escherichia coli was (15.18±0.30) mm. When GPG (100 μmol per mouse) was injected, the life prolongation period of the lethal dose of Escherichia coli was 63.2 h. The above design can be time-saving and labor-saving, so that the target peptide (GPG) can be obtained, with low hemolyticity, strong anticancer and antibacterial activities, and long half-life. The result indicates that GPG is a whole new polypeptide series with clinical application prospect, and GRGDSP chain could be used as N terminal domain of modified melittin and its biological activities and half-life are improved.

Key words: modified melittin, long-effective peptide, molecular design, antitumor activity, antibacterial activity

CLC Number:

S896.5

Ye Ruobai, Wu Zhenhong, Miao Xiaoqing. Design and Bioactivity Evaluation of Long-effective Modified Melittin (GPG) with Antitumor and Antibacterial Function[J]. Chinese Agricultural Science Bulletin, 2020, 36(16): 34-41.

Figures/Tables 6

References 30

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设计序列号	设计多肽的氨基酸序列	简记	整体结构参数
设计序列号	设计多肽的氨基酸序列	简记	净正电荷数	疏水力矩	疏水性	α-螺旋度	两亲性
1	RGDSFLHLAKKFGKAFPAVLKVLTTG		+4	0.420	0.320	50.0	是
2	GRGDSPFLHLAKKFGKAFPAVLKVLTTG		+4	0.557	0.596	66.7	是
3	GRGDSPSFLHLAKKPGKAFPAVLKVLTTG		+4	0.572	0.526	61.1	是
4	RGDFLHLAKKFGKAFPAVLKVLTTG		+4	0.557	0.596	66.7	是
5	GRGDSPKFLHSAKKFGKAFPAVLKVLTTG	GPG	+5	0.565	0.377	58.0	是
6	GRGDSPKFLHLAKKFGKAFIGIAVLKVLTTG		+5	0.451	0.594	38.9	是
7	GRGDSPKFLHSAKKFGKAFIGIAVLKVLTTG		+5	0.439	0.497	44.4	是
8	RGDKFLHLAKKFGKAFIGIAVLKVLTTG		+5	0.451	0.594	38.9	是
9	RGDKFLHSAKKFGKAFIGIAVLKVLTTG		+5	0.439	0.497	55.6	是
10	RGDKFLHLAKKFGKAFPAVLKVLTTG		+5	0.589	0.473	38.9	是
11	GRGDSPKFLHLAKKFGKAFPAVLKVLTTG		+5	0.589	0.473	38.9	是
12	GRGDSPKWKLFKKIPAVLKVLTTG		+5	0.395	0.554	44.4	是
13	RGDKWKLFKKIEKVGQPAVLKVLTTGL		+5	0.283	0.328	50.0	是
14	GRGDSPKWKLFKKIEKVGQPAVLKVLTTGL		+5	0.283	0.328	50.0	是
15	GRGDSPKFLHSAKKFKAFMMEAVLKVLTT		+5	0.239	0.528	61.1	是
16	RGDKFLHSAKKFKAFMMEAVLKVLTT		+5	0.214	0.432	44.4	是
17	RGDKWKLFKKIPKFLHLAKKF	RP18	+7	0.333	0.332	52.4	是
18	RGDKWKLFKKIIGIKFLHSAKKF		+7	0.202	0.505	50.0	是
19	GRGDSPKWKLFKKIIGIKFLHLAKKF		+7	0.298	0.602	44.4	是
20	RGDKFLHSAKKFGKAFPAVLKVLTTG	RPG	+5	0.392	0.448	50.0	是
21	RGDKWKLFKKIPAVLKVLTTG	RCAG	+5	0.395	0.554	44.4	是

设计序列号	设计多肽的氨基酸序列	简记	整体结构参数
设计序列号	设计多肽的氨基酸序列	简记	净正电荷数	疏水力矩	疏水性	α-螺旋度	两亲性
1	RGDSFLHLAKKFGKAFPAVLKVLTTG		+4	0.420	0.320	50.0	是
2	GRGDSPFLHLAKKFGKAFPAVLKVLTTG		+4	0.557	0.596	66.7	是
3	GRGDSPSFLHLAKKPGKAFPAVLKVLTTG		+4	0.572	0.526	61.1	是
4	RGDFLHLAKKFGKAFPAVLKVLTTG		+4	0.557	0.596	66.7	是
5	GRGDSPKFLHSAKKFGKAFPAVLKVLTTG	GPG	+5	0.565	0.377	58.0	是
6	GRGDSPKFLHLAKKFGKAFIGIAVLKVLTTG		+5	0.451	0.594	38.9	是
7	GRGDSPKFLHSAKKFGKAFIGIAVLKVLTTG		+5	0.439	0.497	44.4	是
8	RGDKFLHLAKKFGKAFIGIAVLKVLTTG		+5	0.451	0.594	38.9	是
9	RGDKFLHSAKKFGKAFIGIAVLKVLTTG		+5	0.439	0.497	55.6	是
10	RGDKFLHLAKKFGKAFPAVLKVLTTG		+5	0.589	0.473	38.9	是
11	GRGDSPKFLHLAKKFGKAFPAVLKVLTTG		+5	0.589	0.473	38.9	是
12	GRGDSPKWKLFKKIPAVLKVLTTG		+5	0.395	0.554	44.4	是
13	RGDKWKLFKKIEKVGQPAVLKVLTTGL		+5	0.283	0.328	50.0	是
14	GRGDSPKWKLFKKIEKVGQPAVLKVLTTGL		+5	0.283	0.328	50.0	是
15	GRGDSPKFLHSAKKFKAFMMEAVLKVLTT		+5	0.239	0.528	61.1	是
16	RGDKFLHSAKKFKAFMMEAVLKVLTT		+5	0.214	0.432	44.4	是
17	RGDKWKLFKKIPKFLHLAKKF	RP18	+7	0.333	0.332	52.4	是
18	RGDKWKLFKKIIGIKFLHSAKKF		+7	0.202	0.505	50.0	是
19	GRGDSPKWKLFKKIIGIKFLHLAKKF		+7	0.298	0.602	44.4	是
20	RGDKFLHSAKKFGKAFPAVLKVLTTG	RPG	+5	0.392	0.448	50.0	是
21	RGDKWKLFKKIPAVLKVLTTG	RCAG	+5	0.395	0.554	44.4	是

多肽浓度	GPG	RPG	RP18	RCAG
150.00 μmol/L	0	0	4.0±0.2	0
75.00 μmol/L	0	0	1.8±0.2	0
37.50 μmol/L	0	0	0.2±0.1	0
18.75 μmol/L	0	0	0	0
9.38 μmol/L	0	0	0	0
4.69 μmol/L	0	0	0	0

多肽浓度	GPG	RPG	RP18	RCAG
150.00 μmol/L	0	0	4.0±0.2	0
75.00 μmol/L	0	0	1.8±0.2	0
37.50 μmol/L	0	0	0.2±0.1	0
18.75 μmol/L	0	0	0	0
9.38 μmol/L	0	0	0	0
4.69 μmol/L	0	0	0	0

多肽浓度	对H₂₂抑癌率				对SMMC-7721抑癌率				对SW-1116抑癌率
多肽浓度	GPG	RPG	RP18	RCAG	GPG	RPG	RP18	RCAG	GPG	RPG	RP18	RCAG
33.3 μmol/L	78.0±0.6	74.0±1.4	71.0±1.4	70.0±1.0	74.0±1.2	72.0±1.3	71.0±1.6	70.0±1.3	74.0±0.5	72.0±1.6	72.0±1.5	71.0±1.6
16.7 μmol/L	56.0±0.9	51.3±1.2	48.5±1.1	48.1±1.1	51.0±1.6	49.6±1.5	47.2±1.5	46.7±0.8	50.0±1.5	48.3±1.2	48.5±1.4	47.9±1.4
8.3 μmol/L	32.0±1.4	26.5±1.4	24.7±1.4	24.4±0.8	25.0±1.3	24.2±1.4	22.3±1.4	21.9±1.4	22.5±1.4	20.6±1.5	22.0±1.4	21.9±1.7
4.2 μmol/L	20.1±1.6	15.4±1.5	12.6±1.0	12.4±1.0	13.5±1.0	11.7±1.2	10.5±1.3	10.3±1.4	10.7±0.6	9.8±0.8	11.1±0.5	11.0±0.8
2.1 μmol/L	3.0±1.1	1.7±0.8	0.9±0.6	0.8±0.4	1.2±0.6	0.8±0.6	0.7±0.4	0.7±0.5	0.8±0.4	0.6±0.3	0.7±0.3	0.7±0.7