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De novo design of a monomeric three-stranded antiparallel β-sheet

Published online by Cambridge University Press:  01 April 1999

EVA DE ALBA
Affiliation:
Instituto de Estructura de la Materia, Consejo Superior de Investigaciones Científicas, Serrano 119, 28006 Madrid, Spain Present address: Laboratory of Biophysical Chemistry, National Heart, Lung and Blood Institute, National Institute of Health, Bethesda, Maryland 20892.
JORGE SANTORO
Affiliation:
Instituto de Estructura de la Materia, Consejo Superior de Investigaciones Científicas, Serrano 119, 28006 Madrid, Spain
MANUEL RICO
Affiliation:
Instituto de Estructura de la Materia, Consejo Superior de Investigaciones Científicas, Serrano 119, 28006 Madrid, Spain
M. ANGELES JIMÉNEZ
Affiliation:
Instituto de Estructura de la Materia, Consejo Superior de Investigaciones Científicas, Serrano 119, 28006 Madrid, Spain
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Abstract

Here we describe the NMR conformational study of a 20-residue linear peptide designed to fold into a monomeric three-stranded antiparallel β-sheet in aqueous solution. Experimental and statistical data on amino acid β-turn and β-sheet propensities, cross-strand side-chain interactions, solubility criteria, and our previous experience with β-hairpins were considered for a rational selection of the peptide sequence. Sedimentation equilibrium measurements and NMR dilution experiments provide evidence that the peptide is monomeric. Analysis of 1H and 13C-NMR parameters of the peptide, in particular NOEs and chemical shifts, and comparison with data obtained for two 12-residue peptides encompassing the N- and C-segments of the designed sequence indicates that the 20-residue peptide folds into the expected conformation. Assuming a two-state model, the exchange kinetics between the β-sheet and the unfolded peptide molecules is in a suitable range to estimate the folding rate on the basis of the NMR linewidths of several resonances. The time constant for the coil-β-sheet transition is of the order of several microseconds in the designed peptide. Future designs based on this peptide system are expected to contribute greatly to our knowledge of the many factors involved in β-sheet formation and stability.

Type
Research Article
Copyright
© 1999 The Protein Society

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