The role of histidine-114 of Ssulfolobus acidocaldarius geranylgeranyl diphosphate synthase in chain-length determination.
Sulfolobus acidocaldarius geranylgeranyl diphosphate synthase yields (all-E)-C(20) prenyl diphosphate as a final product. The three-dimensional model of the enzyme suggested that removing two bulky residues at 77 and 114 would allow additional prenyl-chain elongation. To test this, we examined several mutants with substitutions at 77 and/or 114. As a result, the mutants, F77G, F77G and H114A, F77G and H114G, H114A, and H114G gave C(30), C(45), C(50), C(30) and C(40) as the main long product, respectively. These observations indicate that histidine-114 plays a crucial role in chain-length determination along with phenylalanine-77.
Hirooka
K
K
Department of Biochemistry and Engineering, Tohoku University, Aoba Aramaki 07, Aoba-ku, Sendai 980-8579, Japan.
Kato
T
T
Matsu-ura
J
J
Hemmi
H
H
Nishino
T
T
D019883:Alkyl and Aryl Transferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019943:Amino Acid Substitution / Q000235:genetics; D002384:Catalysis; D002855:Chromatography, Thin Layer; D011756:Diphosphates / Q000737:chemistry* / Q000378:metabolism*; D004795:Enzyme Stability; D051231:Farnesyltranstransferase; D006639:Histidine / Q000235:genetics / Q000378:metabolism*; D006358:Hot Temperature; D008958:Models, Molecular; D008970:Molecular Weight; D009154:Mutation / Q000235:genetics; D010649:Phenylalanine / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D013379:Substrate Specificity; D016945:Sulfolobus acidocaldarius / Q000201:enzymology* / Q000235:genetics
D011756:Diphosphates; D010649:Phenylalanine; D006639:Histidine; D019883:Alkyl and Aryl Transferases; D051231:Farnesyltranstransferase