title: Architecture and characterization of sarcosine oxidase from Thermococcus kodakarensis KOD1.
abstract: Sarcosine oxidase (SOX) catalyzes the oxidation of the methyl group in sarcosine and transfer of the oxidized methyl group into the one-carbon metabolic pool. Here, we separately cloned and expressed α and β subunit of SOX from Thermococcus kodakarensis KOD1 (TkSOX) in Escherichia coli and the recombinant proteins were purified to homogeneity. Gel filtration chromatography and transmission electron microscopy analysis showed that the α subunit formed a dimeric structure and behaved as an NADH dehydrogenase; β subunit was a tetramer that had sarcosine oxidase and L: -proline dehydrogenase activity. The TkSOX complex assembled into the hetero-octameric (αβ)(4) form and had NADH dehydrogenase activity. Gold-label analysis indicated that α and β subunits were oriented in the alternative form. Based on these results, we suggested that TkSOX was a multifunctional enzyme and that each subunit and (αβ)(4) complex may separately exist as a function enzyme in different conditions.
authors.affiliation: Division of Applied Life Sciences (BK21 Program), Gyeongsang National University, Jinju, Korea.
authors.forename: Sangmin
authors.initials: S
authors.lastname: Lee
authors.forename: Baolei
authors.initials: B
authors.lastname: Jia
authors.forename: Bang Phuong
authors.initials: BP
authors.lastname: Pham
authors.forename: Yongqi
authors.initials: Y
authors.lastname: Shao
authors.forename: Jae Myeong
authors.initials: JM
authors.lastname: Kwak
authors.forename: Gang-Won
authors.initials: GW
authors.lastname: Cheong
mesh_terms: D001483:Base Sequence; D055162:Biocatalysis; D002850:Chromatography, Gel; D017931:DNA Primers; D008854:Microscopy, Electron; D016133:Polymerase Chain Reaction; D050879:Sarcosine Oxidase / Q000737:chemistry / Q000378:metabolism*; D019712:Thermococcus / Q000201:enzymology*
chemical_list: D017931:DNA Primers; D050879:Sarcosine Oxidase
references.citation: FEBS J. 2005 Aug;272(16):4044-54
references.pmid: 16098188
references.citation: Protein Expr Purif. 2005 Sep;43(1):33-43
references.pmid: 15922624
references.citation: J Biol Chem. 2004 Apr 23;279(17):16947-53
references.pmid: 14766747
references.citation: Genome Res. 2005 Mar;15(3):352-63
references.pmid: 15710748
references.citation: Appl Microbiol Biotechnol. 2006 Dec;73(3):559-66
references.pmid: 16977470
references.citation: Biochim Biophys Acta. 1997 Mar 20;1351(1-2):157-67
references.pmid: 9116029
references.citation: Biochemistry. 1995 Dec 26;34(51):16703-7
references.pmid: 8527444
references.citation: Appl Environ Microbiol. 2001 Apr;67(4):1470-5
references.pmid: 11282592
references.citation: Ultramicroscopy. 1981;6(2):187-94
references.pmid: 7268930
references.citation: J Struct Biol. 1996 Oct;116(1):30-4
references.pmid: 8812976
references.citation: FEBS Lett. 1994 Mar 21;341(2-3):152-5
references.pmid: 7907992
references.citation: Arch Biochem Biophys. 1997 Jun 1;342(1):176-81
references.pmid: 9185627
references.citation: Amino Acids. 1994 Feb;7(1):27-43
references.pmid: 24185971
references.citation: J Biochem. 1981 Feb;89(2):599-607
references.pmid: 7240129
references.citation: Extremophiles. 2004 Apr;8(2):99-108
references.pmid: 15064976
references.citation: J Mol Biol. 2006 Jul 28;360(5):1000-18
references.pmid: 16820168
references.citation: J Biol Chem. 2005 Sep 2;280(35):31045-9
references.pmid: 16027125
references.citation: FEBS Lett. 1988 Dec 5;241(1-2):239-45
references.pmid: 2461878
references.citation: Histochem Cell Biol. 2000 Jun;113(6):489-95
references.pmid: 10933225