Dynamic structure of a highly ordered beta-sheet molten globule: multiple conformations with a stable core

Biochemistry
E BarbarC Woodward

Abstract

The structure of [14-38]Abu, a variant of bovine pancreatic trypsin inhibitor (BPTI) with only the 14-38 disulfide bridge intact, has been analyzed by two-dimensional 1H and 1H-15N NMR. Except for the 18-24, 29-35 antiparallel beta-sheet, residues in all regions of the molecule give two exchange cross peaks for each 1H; for one residue, Gly 37, three exchange cross peaks are observed. The presence of exchange cross peaks indicates that the residues sample conformations that interconvert on a time scale of milliseconds or longer. Over 90% of the NMR spectra have been assigned, including backbone and side chain atoms and their exchange cross peaks. Analyses of chemical shifts, chemical exchange, hydrogen isotope exchange, and NOEs indicate that [14-38]Abu at pH 4.5 and 1 degree C is an ensemble of interconverting conformations, in all of which the 18-24, 29-35 antiparallel beta-sheet is native-like and intact. Outside the antiparallel beta-sheet, residues undergo local order/disorder transitions. The stable structure of [14-38]Abu is not in the vicinity of the 14-38 disulfide bond but rather is in the slow-exchange core. NOE analysis indicates that the main tertiary interactions involve hydrophobic contacts with the rings of Tyr ...Continue Reading

Citations

Mar 19, 1999·Protein Science : a Publication of the Protein Society·K J BarnhamR S Norton
Dec 19, 2002·Biophysical Chemistry·Natàlia CarullaClare Woodward
Feb 28, 1997·Biophysical Chemistry·E BarbarC Woodward
Feb 1, 1996·Current Opinion in Structural Biology·D R Shortle
Feb 1, 1996·Current Opinion in Structural Biology·A D Miranker, C M Dobson
May 31, 2001·Journal of Molecular Graphics & Modelling·C WoodwardG Barany
May 22, 2002·Protein Science : a Publication of the Protein Society·Natàlia CarullaGeorge Barany
Aug 19, 1999·Protein Science : a Publication of the Protein Society·R Li, C Woodward
Sep 24, 1999·Protein Science : a Publication of the Protein Society·G Bulaj, D P Goldenberg
Aug 1, 1997·Biological Chemistry·T E Creighton

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