title: Functional and structural characteristics of methylmalonyl-CoA mutase from Pyrococcus horikoshii.
abstract: Methylmalonyl-CoA mutase (MCM) requires 5'-deoxyadenosylcobalamin (AdoCbl) as a cofactor and is widely distributed in organisms from bacteria and animals. Although genes encoding putative MCMs are present in many archaea, they are separately encoded in large and small subunits. The large and small subunits of archaeal MCM are similar to the catalytic and AdoCbl-binding domains of human MCM, respectively. In Pyrococcus horikoshii OT3, putative genes PH1306 and PH0275 encode the large and small subunits, respectively. Because information on archaeal MCM is extremely restricted, we examined the functional and structural characteristics of P. horikoshii MCM. Reconstitution experiments using recombinant PH0275 and PH1306 showed that these proteins assemble in equimolar ratios and form of heterotetrameric complexes in the presence of AdoCbl. Subsequent immunoprecipitation experiments using anti-PH0275 and anti-PH1306 antibodies suggested that PH0275 and PH1306 form a complex in P. horikoshii cells in the presence of AdoCbl.
authors.affiliation: a Faculty of Agriculture, School of Agricultural, Biological, and Environmental Sciences , Tottori University , Tottori , Japan.
authors.forename: Yukinori
authors.initials: Y
authors.lastname: Yabuta
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Kamei
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Bito
authors.forename: Jiro
authors.initials: J
authors.lastname: Arima
authors.forename: Kazunari
authors.initials: K
authors.lastname: Yoneda
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Sakuraba
authors.forename: Toshihisa
authors.initials: T
authors.lastname: Ohshima
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Nakano
authors.forename: Fumio
authors.initials: F
authors.lastname: Watanabe
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003038:Cobamides / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D008765:Methylmalonyl-CoA Mutase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D055503:Protein Multimerization; D043902:Pyrococcus horikoshii / Q000201:enzymology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D003038:Cobamides; D011994:Recombinant Proteins; D008765:Methylmalonyl-CoA Mutase; C000913:cobamamide
keywords: 5′-deoxyadenosylcobalamin; Pyrococcus horikoshii; archaea; methylmalonyl-CoA mutase; vitamin B12