s. aureus mscl is a pentamer in vivo but of variable stoichiometries in vitro implications for detergent-solubilized membrane proteins金黄色葡萄球菌mscl体内五聚物但变量化学计量学在体外对detergent-solubilized膜蛋白的影响.pdfVIP

S. aureus MscL Is a Pentamer In Vivo but of Variable Stoichiometries In Vitro: Implications for Detergent- Solubilized Membrane Proteins 1,2 1 3 1,2 1 Michael R. Dorwart , Robin Wray , Chad A. Brautigam , Youxing Jiang , Paul Blount * 1 Department of Physiology, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas, United States of America, 2 Howard Hughes Medical Institute, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas, United States of America, 3 Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas, United States of America Abstract While the bacterial mechanosensitive channel of large conductance (MscL) is the best studied biological mechanosensor and serves as a paradigm for how a protein can sense and respond to membrane tension, the simple matter of its oligomeric state has led to debate, with models ranging from tetramers to hexamers. Indeed, two different oligomeric states of the bacterial mechanosensitive channel MscL have been resolved by X-ray crystallography: The M. tuberculosis channel (MtMscL) is a pentamer, while the S. aureus protein (SaMscL) forms a tetramer. Because several studies suggest that, like MtMscL, the E. coli MscL (EcoMscL) is a pentamer, we re-investigated the oligomeric state of SaMscL. To determine the structural organization of MscL in the cell membrane we developed a disulfide-trapping approach. Surprisingly, we found that virtually all SaMscL channels in vivo are pentameric, indicating this as the physiologically relevant and functional oligomeric state. Complementing our in vivo results, we