Integral Membrane Proteins in Nanodiscs Can Be Studied by Solution NMR Spectroscopy

详细信息    查看全文

• 作者：Julian M. Glck ; Marc Wittlich ; Sophie Feuerstein ; Silke Hoffmann ; Dieter Willbold ; Bernd W. Koenig
• 刊名：Journal of the American Chemical Society
• 出版年：2009
• 出版时间：September 2, 2009
• 年：2009
• 卷：131
• 期：34
• 页码：12060-12061
• 全文大小：158K
• 年卷期：v.131,no.34(September 2, 2009)
• ISSN：1520-5126

文摘

We present a two-dimensional solution NMR spectrum of an integral membrane protein (IMP) in a nanodisc. Solution NMR relies on rapid isotropic tumbling of the analyte with correlation times in the nanosecond range. IMPs in a cellular membrane do not satisfy this condition. Previous liquid-state NMR studies on IMPs were conducted in organic solvent or artificial membrane mimicking particles like detergent micelles. Nanodiscs are relatively small (150 kDa), detergent-free model membranes that are suitable for functional reconstitution of IMPs. Nanodiscs allow solubilization of integral membrane proteins in a nearly native lipid bilayer environment. The 70 residue polypeptide CD4mut was incorporated into nanodiscs. CD4mut features one transmembrane helix. The aliphatic ¹H−¹³C HSQC spectrum of nanodiscs with inserted, (¹³C, ¹⁵N)-labeled CD4mut exhibits reasonably dispersed protein and lipid NMR signals. Our results demonstrate that IMPs in nanodiscs are amenable to liquid-state NMR methodology.