Apoptin NLS2 homodimerization strategy for improved antibacterial activity and bio-stability

IF 3 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Amino Acids Pub Date : 2023-09-19 DOI:10.1007/s00726-023-03321-1

Anu Kumari, Mahavir Singh, Ruchi Sharma, Tarun Kumar, Naresh Jindal, Sushila Maan, Vinay G. Joshi

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Abstract

The emergence of antibiotic resistance prompts exploration of viable antimicrobial peptides (AMPs) designs. The present study explores the antimicrobial prospects of Apoptin nuclear localization sequence (NLS2)-derived peptide ANLP (PRPRTAKRRIRL). Further, we examined the utility of the NLS dimerization strategy for improvement in antimicrobial activity and sustained bio-stability of AMPs. Initially, the antimicrobial potential of ANLP using antimicrobial peptide databases was analyzed. Then, ANLP along with its two homodimer variants namely ANLP-K1 and ANLP-K2 were synthesized and evaluated for antimicrobial activity against Escherichia coli and Salmonella. Among three AMPs, ANLP-K2 showed efficient antibacterial activity with 12 µM minimum inhibitory concentration (MIC). Slow degradation of ANLP-K1 (26.48%) and ANLP-K2 (13.21%) compared with linear ANLP (52.33%) at 480 min in serum stability assay indicates improved bio-stability of dimeric peptides. The AMPs presented no cytotoxicity in Vero cells. Dye penetration assays confirmed the membrane interacting nature of AMPs. The zeta potential analysis reveals effective charge neutralization of both lipopolysaccharide (LPS) and bacterial cells by dimeric AMPs. The dimeric AMPs on scanning electron microscopy studies showed multiple pore formations on the bacterial surface. Collectively, proposed Lysine scaffold dimerization of Apoptin NLS2 strategy resulted in enhancing antibacterial activity, bio-stability, and could be effective in neutralizing the off-target effect of LPS. In conclusion, these results suggest that nuclear localization sequence with a modified dimeric approach could represent a rich source of template for designing future antimicrobial peptides.

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Apoptin NLS2同源二聚策略可提高抗菌活性和生物稳定性。

抗生素耐药性的出现促使人们探索可行的抗菌肽（AMP）设计。本研究探讨了Apoptin核定位序列（NLS2）衍生肽ANLP（PRRTAKRRIRL）的抗菌前景。此外，我们研究了NLS二聚化策略在提高抗菌活性和持续生物稳定性方面的效用。最初，使用抗菌肽数据库分析ANLP的抗菌潜力。然后，合成了ANLP及其两个同源二聚体变体，即ANLP-K1和ANLP-K2，并评估了其对大肠杆菌和沙门氏菌的抗菌活性。在三种AMP中，ANLP-K2表现出有效的抗菌活性，最小抑菌浓度为12µM。在血清稳定性测定中，与线性ANLP（52.33%）相比，ANLP-K1（26.48%）和ANLP-K2（13.21%）在480分钟时的缓慢降解表明二聚肽的生物稳定性提高。AMPs在Vero细胞中没有表现出细胞毒性。染料渗透分析证实了AMPs的膜相互作用性质。ζ电位分析揭示了二聚体AMP对脂多糖（LPS）和细菌细胞的有效电荷中和作用。扫描电子显微镜研究的二聚体AMPs显示细菌表面形成了多个孔。总之，所提出的赖氨酸支架二聚Apoptin NLS2策略提高了抗菌活性和生物稳定性，并可有效中和LPS的脱靶作用。总之，这些结果表明，采用改良二聚体方法的核定位序列可以为设计未来的抗菌肽提供丰富的模板来源。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Amino Acids 生物-生化与分子生物学

CiteScore

6.40

自引率

5.70%

发文量

审稿时长

2.2 months

期刊介绍： Amino Acids publishes contributions from all fields of amino acid and protein research: analysis, separation, synthesis, biosynthesis, cross linking amino acids, racemization/enantiomers, modification of amino acids as phosphorylation, methylation, acetylation, glycosylation and nonenzymatic glycosylation, new roles for amino acids in physiology and pathophysiology, biology, amino acid analogues and derivatives, polyamines, radiated amino acids, peptides, stable isotopes and isotopes of amino acids. Applications in medicine, food chemistry, nutrition, gastroenterology, nephrology, neurochemistry, pharmacology, excitatory amino acids are just some of the topics covered. Fields of interest include: Biochemistry, food chemistry, nutrition, neurology, psychiatry, pharmacology, nephrology, gastroenterology, microbiology