title: Structural characterization of the zinc site in protein farnesyltransferase.
abstract: X-ray absorption spectroscopy has been used to determine the structure of the Zn site in protein farnesyltransferase. Extended X-ray absorption fine structure (EXAFS) data are consistent with a Zn site that is ligated to three low-Z (oxygen or nitrogen) ligands and one cysteine sulfur, as predicted from the crystal structures that are available for farnesyltransferase. However, in contrast with the crystallographic results the EXAFS data do not show evidence for significant distortions in the Zn-ligand distances. The average Zn-(N/O) and Zn-S distances are 2.04 and 2.31 A, respectively. Addition of a farnesyl diphosphate analogue causes no detectable change in the structure of the Zn site. However, addition of peptide substrate causes a change in ligation from ZnS(N/O)(3) to ZnS(2)(N/O)(2), consistent with ligation of the C-terminal cysteine to the Zn. There is no significant change in Zn-ligand distances when a substrate binds, demonstrating that the Zn remains four-coordinate. Addition of both peptide and farnesyl diphosphate to give the product complex causes the Zn to return to ZnS(N/O)(3) ligation, indicating that the product thioether is not tightly coordinated to the Zn. These spectroscopic experiments provide insight into the catalytic mechanism of FTase.
authors.affiliation: Department of Chemistry and Biophysics Research Division, The University of Michigan, Ann Arbor, MH 48109-1055, USA.
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Tobin
authors.forename: Jennifer S
authors.initials: JS
authors.lastname: Pickett
authors.forename: Heather L
authors.initials: HL
authors.lastname: Hartman
authors.forename: Carol A
authors.initials: CA
authors.lastname: Fierke
authors.forename: James E
authors.initials: JE
authors.lastname: Penner-Hahn
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray; D051231:Farnesyltranstransferase; D007700:Kinetics; D013057:Spectrum Analysis / Q000379:methods; D015032:Zinc / Q000737:chemistry*
chemical_list: D019883:Alkyl and Aryl Transferases; D051231:Farnesyltranstransferase; D015032:Zinc