Observation of sequential steps in the folding of intestinal fatty acid binding protein using a slow folding mutant and 19F NMR.

ABSTRACT: The rat intestinal fatty acid binding protein (IFABP) primarily comprises two beta-sheet structures surrounding a large internal cavity. The urea denatured WT protein folds within seconds after dilution to nondenaturing conditions. Replacing a glycine with valine in the turn between the last two beta-strands (Gly121Val) slows the folding process by more than three orders of magnitude. After incorporating 4-(19)F-phenylalanine into the mutant protein, we were able to directly monitor the behavior of the eight phenylalanine side chains in real time during folding using (19)F NMR. Specifically, there is a nonnative-like collapse in regions involving three phenylalanine residues (Phe-62, Phe-68, and Phe-93) within milliseconds. At least two distinct NMR peaks were observed, suggesting conforma

SUBMITTER: Li H 

PROVIDER: S-EPMC1924545 | biostudies-literature | 2007 Jul

REPOSITORIES: biostudies-literature