rational mutational analysis of a multidrug mfs transporter camdr1p of candida albicans by employing a membrane environment based computational approach理性突变分析多种mfs运输车camdr1p白色念珠菌采用膜环境为基础的计算方法.pdfVIP

Rational Mutational Analysis of a Multidrug MFS Transporter CaMdr1p of Candida albicans by Employing a Membrane Environment Based Computational Approach 1. 2. 2 1¤ 2 1 Khyati Kapoor , Mohd Rehan , Ajeeta Kaushiki , Ritu Pasrija , Andrew M. Lynn *, Rajendra Prasad * 1 School of Life Sciences, Jawaharlal Nehru University, New Delhi, India, 2 School of Information Technology, Jawaharlal Nehru University, New Delhi, India Abstract CaMdr1p is a multidrug MFS transporter of pathogenic Candida albicans. An over-expression of the gene encoding this protein is linked to clinically encountered azole resistance. In-depth knowledge of the structure and function of CaMdr1p is necessary for an effective design of modulators or inhibitors of this efflux transporter. Towards this goal, in this study, we have employed a membrane environment based computational approach to predict the functionally critical residues of CaMdr1p. For this, information theoretic scores which are variants of Relative Entropy (Modified Relative Entropy REM) were calculated from Multiple Sequence Alignment (MSA) by separately considering distinct physico-chemical properties of transmembrane (TM) and inter-TM regions. The residues of CaMdr1p with high REM which were predicted to be significantly important were subjected to site-directed mutational analysis. Interestingly, heterologous host Saccharomyces cerevisiae, over-expressing these mutant variants of CaMdr1p wherein these high REM residues were replaced by either alanine or leucine, demonstrated increased susceptibility to tested drugs. The hypersensitivity to drugs was supported by abrogated substrate efflux medi