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Science 31 August 1990:
Vol. 249. no. 4972, pp. 1012 - 1016
DOI: 10.1126/science.2204109
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Articles

Science, Vol 249, Issue 4972, 1012-1016
Copyright © 1990 by American Association for the Advancement of Science

articles

Regulation of an enzyme by phosphorylation at the active site

JH Hurley, AM Dean, JL Sohl, DE Koshland Jr, and RM Stroud

Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0448.

The isocitrate dehydrogenase of Escherichia coli is an example of a ubiquitous class of enzymes that are regulated by covalent modification. In the three-dimensional structure of the enzyme-substrate complex, isocitrate forms a hydrogen bond with Ser113, the site of regulatory phosphorylation. The structures of Asp113 and Glu113 mutants, which mimic the inactivation of the enzyme by phosphorylation, show minimal conformational changes from wild type, as in the phosphorylated enzyme. Calculations based on observed structures suggest that the change in electrostatic potential when a negative charge is introduced either by phosporylation or site-directed mutagenesis is sufficient to inactivate the enzyme. Thus, direct interaction at a ligand binding site is an alternative mechanism to induced conformational changes from an allosteric site in the regulation of protein activity by phosphorylation.


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