Folding of a β-sheet Protein Monitored by Real-time NMR Spectroscopy

At low ionic strength, apoplastocyanin forms an unfolded state under non-denaturing conditions. The refolding of this state is sufficiently slow to allow real-time NMR experiments to be performed. Folding of apoplastocyanin, initiated by the addition of salt and followed by real-time 2D 1H– 15N hete...

Full description

Saved in:
Bibliographic Details
Published in:Journal of molecular biology 2003-05, Vol.328 (5), p.1161-1171
Main Authors: Mizuguchi, Mineyuki, Kroon, Gerard J, Wright, Peter E, Jane Dyson, H
Format: Article
Language:English
Subjects:
Apoproteins - chemistry
In Vitro Techniques
Kinetics
Models, Molecular
Nuclear Magnetic Resonance, Biomolecular
plastocyanin
Plastocyanin - chemistry
proline isomerism
Protein Folding
Protein Structure, Secondary
Recombinant Proteins - chemistry
slow protein folding
β-sheet folding
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:At low ionic strength, apoplastocyanin forms an unfolded state under non-denaturing conditions. The refolding of this state is sufficiently slow to allow real-time NMR experiments to be performed. Folding of apoplastocyanin, initiated by the addition of salt and followed by real-time 2D 1H– 15N heteronuclear single quantum coherence (HSQC) spectroscopy, is highly cooperative. A concomitant increase in the intensity of both sequential and long-range nuclear Overhauser effects (NOEs) between backbone amide protons in successive acquisitions of 1H– 15N HSQC-NOESY-HSQC spectra provides the first direct observation of the development of structure-specific NOEs as a protein folds. Our results show that the local and long-range interactions in the native apoplastocyanin are formed simultaneously, consistent with highly cooperative formation of the native structure.
ISSN:0022-2836
1089-8638
DOI:10.1016/S0022-2836(03)00349-8