NMR characterization of HtpG, the E. coli Hsp90, using sparse labeling with ¹³C-methyl alanine.

ABSTRACT: A strategy for acquiring structural information from sparsely isotopically labeled large proteins is illustrated with an application to the E. coli heat-shock protein, HtpG (high temperature protein G), a 145 kDa dimer. It uses ¹³C-alanine methyl labeling in a perdeuterated background to take advantage of the sensitivity and resolution of Methyl-TROSY spectra, as well as the backbone-centered structural information from ¹H-¹³C residual dipolar couplings (RDCs) of alanine methyl groups. In all, 40 of the 47 expected crosspeaks were resolved and 36 gave RDC data. Assignments of crosspeaks were partially achieved by transferring assignments from those made on individual domains using triple resonance methods. However, these were incomplete and in many cases th

SUBMITTER: Pederson K 

PROVIDER: S-EPMC5546222 | biostudies-literature | 2017 Jul

REPOSITORIES: biostudies-literature