College of Biological Engineering, Henan University of Technology, 450001, Zhengzhou, Henan, PR China; Key Laboratory of Functional Molecules for Biomedical Research, Henan University of Technology, 450001, Zhengzhou, Henan, PR China.
College of Biological Engineering, Henan University of Technology, 450001, Zhengzhou, Henan, PR China; Key Laboratory of Functional Molecules for Biomedical Research, Henan University of Technology, 450001, Zhengzhou, Henan, PR China.
Biochem Biophys Res Commun. 2023 Mar 15;649:101-109. doi: 10.1016/j.bbrc.2023.01.083. Epub 2023 Feb 1.
Candida tropicalis is often reported as the second or third most common pathogen causing fungal infections. Antimicrobial peptides (AMPs) have attracted increasing attention for their broad-spectrum antimicrobial properties and low cytotoxicity. Our previous studies have shown that CGA-N9, a non-membrane-rupturing AMP, crosses the cell membrane to exert anticandidal activity. We speculate that there are some related transporters that assist in the transmembrane transport of CGA-N9. In this study, the relationship between CGA-N9 lethality kinetics and its real-time transmembrane amount in C. tropicalis cells was investigated. The results demonstrated that there was a positive correlation between its candicidal activity and transmembrane amount. A total of 12 oligopeptide transporter (OPT) coding sequences (CDSs) were cloned from C. tropicalis by using the conservative OPT gene sequences of Candida spp. to design primers and were named C. tropicalis OPTs (CtOPTs). The results of RT‒qPCR demonstrated that the expression levels of CtOPT1, CtOPT9 and CtOPT12 were correlated with the CGA-N9 transmembrane amount in a time-dependent manner. The results of molecular docking demonstrated that CtOPT1, CtOPT9 and CtOPT12 interact strongly with CGA-N9. Therefore, CtOPT1, CtOPT9 and CtOPT12 were predicted to assist in the transmembrane transport of the AMP CGA-N9.
热带假丝酵母通常被报道为引起真菌感染的第二或第三大常见病原体。抗菌肽 (AMP) 因其广谱抗菌特性和低细胞毒性而受到越来越多的关注。我们之前的研究表明，CGA-N9 是一种非破坏细胞膜的 AMP，它穿过细胞膜发挥抗真菌活性。我们推测有一些相关的转运蛋白有助于 CGA-N9 的跨膜转运。在这项研究中，研究了 CGA-N9 致死动力学与其在热带假丝酵母细胞中的实时跨膜量之间的关系。结果表明，其杀真菌活性与其跨膜量之间存在正相关关系。通过使用念珠菌属的保守 OPT 基因序列设计引物，从热带假丝酵母中克隆了 12 个寡肽转运蛋白 (OPT) 编码序列 (CDS)，并将其命名为热带假丝酵母 OPT (CtOPT)。RT‒qPCR 的结果表明，CtOPT1、CtOPT9 和 CtOPT12 的表达水平与 CGA-N9 的跨膜量呈时间依赖性相关。分子对接的结果表明，CtOPT1、CtOPT9 和 CtOPT12 与 CGA-N9 强烈相互作用。因此，预测 CtOPT1、CtOPT9 和 CtOPT12 有助于 AMP CGA-N9 的跨膜转运。
