Improved strategy for isoleucine ¹H/¹³C methyl labeling in Pichia pastoris.

ABSTRACT: Site specific methyl labeling combined with methyl TROSY offers a powerful NMR approach to study structure and dynamics of proteins and protein complexes of high molecular weight. Robust and cost-effective methods have been developed for site specific protein ¹H/¹³C methyl labeling in an otherwise deuterated background in bacteria. However, bacterial systems are not suitable for expression and isotope labeling of many eukaryotic and membrane proteins. The yeast Pichia pastoris (P. pastoris) is a commonly used host for expression of eukaryotic proteins, and site-specific methyl labeling of perdeuterated eukaryotic proteins has recently been achieved with this system. However, the practical utility of methyl labeling and deuteration in P. pastoris is limited by high cos

SUBMITTER: Ali R 

PROVIDER: S-EPMC6875547 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature