Amide exchange rates in Escherichia coli acyl carrier protein: correlation with protein structure and dynamics.
The acyl carrier protein (ACP) of Escherichia coli is a 77-amino acid, highly negatively charged three-helix protein that plays a central role in fatty acid biosynthesis. Previous NMR studies have suggested the presence of multiple conformations and marginally stable secondary structural elements. The stability of these elements is now examined by monitoring amide exchange in apo-ACP using NMR-based methods. Because ACP exhibits many rapid exchange rates, application of traditional isotope exchange methods is difficult. In one approach, heteronuclear correlation experiments with pulsed field-gradient coherence selection have reduced the time needed to collect two-dimensional 1H-15N correlation spectra to the point where measurement of exchange of amide protons for deuterium on the timescale of minutes can be made. In another approach, water proton selective inversion-exchange experiments were performed to estimate the exchange rates of protons exchanging on timescales of less than a second. Backbone amide protons in the region of helix II were found to exchange significantly more rapidly than those in helices I and III, consistent with earlier structural models suggesting a dynamic disruption of the second helix. Highly protected amides occur on faces of the helices that may pack into a hydrophobic core present in a partially disrupted state.
Andrec
M
M
Department of Chemistry, Yale University, New Haven, Connecticut 06511, USA.
Hill
R B
RB
Prestegard
J H
JH
D000213:Acyl Carrier Protein / Q000737:chemistry* / Q000378:metabolism; D000577:Amides / Q000378:metabolism*; D003903:Deuterium / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry; D006859:Hydrogen / Q000378:metabolism; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008433:Mathematics; D011487:Protein Conformation; D017433:Protein Structure, Secondary
D000213:Acyl Carrier Protein; D000577:Amides; D006859:Hydrogen; D003903:Deuterium