Delineation of the allosteric mechanism of a cytidylyltransferase exhibiting negative cooperativity.
The dimeric enzyme CTP:glycerol-3-phosphate cytidylyltransferase (GCT) displays strong negative cooperativity between the first and second binding of its substrate, CTP. Using NMR to study the allosteric mechanism of this enzyme, we observe widespread chemical shift changes for the individual CTP binding steps. Mapping these changes onto the molecular structure allowed the formulation of a detailed model of allosteric conformational change. Upon the second step of ligand binding, NMR experiments indicate an extensive loss of conformational exchange broadening of the backbone resonances of GCT. This suggests that a fraction of the free energy of negative cooperativity is entropic in origin.
Stevens
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Biophysics Research Division, University of Michigan, 930 North University, Ann Arbor, Michigan 48109-1055, USA.
Sanker
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Kent
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Zuiderweg
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D000494:Allosteric Regulation; D000495:Allosteric Site; D019943:Amino Acid Substitution; D001120:Arginine / Q000378:metabolism; D003570:Cytidine Triphosphate / Q000378:metabolism; D019281:Dimerization; D019277:Entropy; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D008954:Models, Biological; D008958:Models, Molecular; D009713:Nucleotidyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D017186:Titrimetry
D008024:Ligands; D003570:Cytidine Triphosphate; D001120:Arginine; D009713:Nucleotidyltransferases; C020675:glycerol-3-phosphate cytidylyltransferase