Sparse C-13 labelling for solid-state NMR studies of P. pastoris expressed eukaryotic seven-transmembrane proteins

作者：Liu, Jing; Liu, Chang; Fan, Ying; Munro, Rachel A.; Ladizhansky, Vladimir; Brown, Leonid S.; Wang, Shenlin^*

来源：Journal of Biomolecular NMR, 2016, 65(1): 7-13.

DOI：10.1007/s10858-016-0033-1

• Magic-angle-spinning solid-state NMR
• Sparse C-13 isotope labelling
• Eukaryotic seven-transmembrane proteins
• P. pastoris expression system

We demonstrate a novel sparse C-13 labelling approach for methylotrophic yeast P. pastoris expression system, towards solid-state NMR studies of eukaryotic membrane proteins. The labelling scheme was achieved by co-utilizing natural abundance methanol and specifically C-13 labelled glycerol as carbon sources in the expression medium. This strategy improves the spectral resolution by 1.5 fold, displays site-specific labelling patterns, and has advantages for collecting long-range distance restraints for structure determination of large eukaryotic membrane proteins by solid-state NMR.

• 出版日期2016-5
• 单位北京大学