Structural role for Tyr-104 in Escherichia coli isopentenyl-diphosphate isomerase: site-directed mutagenesis, enzymology, and protein crystallography

The Journal of Biological Chemistry
Jérôme de RuyckJohan Wouters

Abstract

Isopentenyl-diphosphate (IPP):dimethylallyl diphosphate isomerase is a key enzyme in the biosynthesis of isoprenoids. The mechanism of the isomerization reaction involves protonation of the unactivated carbon-carbon double bond in the substrate, but identity of the acidic moiety providing the proton is still not clear. Multiple sequence alignments and geometrical features observed in crystal structures of complexes with IPP isomerase suggest that Tyr-104 could play an important role during catalysis. A series of mutants was constructed by directed mutagenesis and characterized by enzymology. Crystallographic and thermal denaturation data for Y104A and Y104F mutants were obtained. Those data demonstrate the importance of residue Tyr-104 for proper folding of Escherichia coli type I IPP isomerase.

References

Jan 1, 1985·Methods in Enzymology·D M Satterwhite
Nov 5, 1997·Science·J C Sacchettini, C D Poulter
Feb 7, 2001·Proceedings of the National Academy of Sciences of the United States of America·K KanedaH Seto
Feb 15, 2001·Acta Crystallographica. Section D, Biological Crystallography·Y OudjamaL Droogmans
Nov 8, 2001·Proceedings of the National Academy of Sciences of the United States of America·J B BonannoS K Burley
Mar 13, 2003·Journal of the American Chemical Society·Johan WoutersEric Oldfield
Sep 17, 2004·Journal of the American Chemical Society·Christina N Carrigan, C Dale Poulter
Jan 1, 1997·Methods in Enzymology·Zbyszek Otwinowski, Wladek Minor

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.