Theoretical studies on farnesyl transferase: evidence for thioether product coordination to the active-site zinc sphere.
Farnesyltransferase (FTase), an interesting zinc metaloenzyme, has been the subject of great attention in anticancer research over the last decade. However, despite the major accomplishments in the field, some very pungent questions on the farnesylation mechanism still persist. In this study, the authors have analyzed a mechanistic paradox that arises from the existence of several contradicting and inconclusive experimental evidence regarding the existence of direct coordination between the active-site zinc cation and the thioether from the farnesylated peptide product, which include UV-vis spectroscopy data on a Co(2+)-substituted FTase, two X-ray crystallographic structures of the FTase-product complex, and extended X-ray absorption fine structure results. Using high-level theoretical calculations on two models of different sizes, and QM/MM calculations on the full enzyme, the authors have shown that the farnesylated product is Zn coordinated, and that a subsequent step where this Zn bond is broken is coherent with the available kinetic results. Furthermore, an explanation for the contradicting experimental evidence is suggested.
Sousa
Sérgio Filipe
SF
REQUIMTE, Departamento de Química, Faculdade de Ciências, Universidade do Porto, Porto, Portugal.
Fernandes
Pedro Alexandrino
PA
Ramos
Maria João
MJ
D001665:Binding Sites; D018360:Crystallography, X-Ray; D051231:Farnesyltranstransferase / Q000737:chemistry* / Q000378:metabolism*; D007477:Ions / Q000737:chemistry; D008958:Models, Molecular; D010100:Oxygen / Q000737:chemistry; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013440:Sulfides / Q000737:chemistry* / Q000378:metabolism*; D015032:Zinc / Q000737:chemistry* / Q000378:metabolism*
D007477:Ions; D010455:Peptides; D013440:Sulfides; D051231:Farnesyltranstransferase; D015032:Zinc; D010100:Oxygen