Bandyopadhyay Susmita, Junjie Ryan Loh, Lim Brendan, Sanjeev R, Xin Woon Yong, Yee Chong Kok, Hui Melodies Sim Ming, Yow Nicole, Sivaraman J, Chatterjee Chiradip
School of Applied Science, Republic Polytechnic, Singapore.
Biopolymers. 2014 Dec;101(12):1143-53. doi: 10.1002/bip.22519.
Ctriporin peptide (Ctr), a novel antimicrobial peptide isolated from the venom of the scorpion Chaerilus tricostatus, shows a broad-spectrum of antimicrobial activity and is able to inhibit antibiotic resistant pathogens, including Methicillin resistant Staphylococcus aureus, Methicillin Resistant Coagulase-negative Staphylococcus, and Penicillin Resistant Staphylococcus epidermidis strains. To understand the active conformation of the Ctr peptide in membranes, we have investigated the interaction of Ctr with the negatively charged and zwitterionic membrane-mimetic micelles such as sodium dodecyl sulphate (SDS) and n-dodecylphosphocholine (DPC), respectively. The interactions were studied using fluorescence and circular dichroism (CD) spectroscopy. Fluorescence experiments revealed that the N-terminus tryptophan residue of Ctr interacted with the hydrophobic core of the membrane mimicking micelles. The CD results suggest that interactions with membrane-mimetic micelles induce an α-helix conformation in Ctr. Moreover, we have determined the solution structures of Ctr in SDS and DPC micelles using nuclear magnetic resonance (NMR) spectroscopy. The structural comparison of Ctr in the presence of SDS and DPC micelles showed significant conformational changes. The observed structural differences of Ctr in anionic versus zwitterionic membrane-mimetic micelles suggest that the mode of interaction of this peptide may be different in two environments which may account for its ability to differentiate bacterial and eukaryotic cell membrane.
Ctriporin肽(Ctr)是从三肋钳蝎毒液中分离出的一种新型抗菌肽,具有广谱抗菌活性,能够抑制耐抗生素病原体,包括耐甲氧西林金黄色葡萄球菌、耐甲氧西林凝固酶阴性葡萄球菌和耐青霉素表皮葡萄球菌菌株。为了了解Ctr肽在膜中的活性构象,我们分别研究了Ctr与带负电荷和两性离子的膜模拟胶束(如十二烷基硫酸钠(SDS)和正十二烷基磷酰胆碱(DPC))的相互作用。使用荧光和圆二色性(CD)光谱研究了这些相互作用。荧光实验表明,Ctr的N端色氨酸残基与膜模拟胶束的疏水核心相互作用。CD结果表明,与膜模拟胶束的相互作用在Ctr中诱导出α-螺旋构象。此外,我们使用核磁共振(NMR)光谱确定了Ctr在SDS和DPC胶束中的溶液结构。Ctr在SDS和DPC胶束存在下的结构比较显示出显著的构象变化。在阴离子型与两性离子型膜模拟胶束中观察到的Ctr结构差异表明,该肽在两种环境中的相互作用模式可能不同,这可能解释了其区分细菌和真核细胞膜的能力。