O-acetylserine sulfhydrylase from Methanosarcina thermophila

被引:26
作者
Borup, B
Ferry, JG
机构
[1] Penn State Univ, Dept Biochem & Mol Biol, University Pk, PA 16802 USA
[2] Penn State Univ, Dept Chem, University Pk, PA 16802 USA
关键词
D O I
10.1128/JB.182.1.45-50.2000
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Cysteine is the major source of fixed sulfur for the synthesis of sulfur-containing compounds in organisms of the Bacteria and Eucarya domains. Though pathways for cysteine biosynthesis have been established for both of these domains, it is unknown how the Archaea fix sulfur or synthesize cysteine, None of the four archaeal genomes sequenced to date contain open reading frames with identities to either O-acetyl-L-serine sulfhydrylase (OASS) or homocysteine synthase, the only sulfur-fixing enzymes known in nature. We report the purification and characterization of OASS from acetate-grown Methanosarcina thermophila, a moderately thermophilic methanoarchaeon. The purified OASS Contained pyridoxal 5'-phosphate and catalyzed the formation of L-cysteine and acetate from O-acetyl-L-serine and sulfide. The N-terminal amino acid sequence has high sequence similarity with other known OASS enzymes from the Eucarya and Bacteria domains. The purified OASS had a specific activity of 129 mu mol of cysteine/min/mg, with a K-m of 500 +/- 80 mu M for sulfide, acid exhibited positive cooperativity and substrate inhibition with O-acetyl-L-serine, Sodium dodecyl sulfate; polyacrylamide gel electrophoresis revealed a single band at 36 kDa, and native gel filtration chromatography indicated a molecular mass of 93 kDa, suggesting that the purified OASS is either a homodimer or a homotrimer, The optimum temperature for activity was between 40 and 60 degrees C, consistent with the optimum growth temperature for M, thermophila, The results of:this study provide the first evidence for a sulfur-fixing enzyme in the Archaea domain. The results also provide the first biochemical evidence for an enzyme with the potential for involvement in cysteine biosynthesis in the Archaea.
引用
收藏
页码:45 / 50
页数:6
相关论文
共 49 条
  • [1] ALTSCHUL SF, 1990, J MOL BIOL, V215, P403, DOI 10.1006/jmbi.1990.9999
  • [2] BECKER MA, 1969, J BIOL CHEM, V244, P2418
  • [3] BENDER DA, 1985, AMINO ACID METABOLIS
  • [4] PURIFICATION AND INITIAL KINETIC CHARACTERIZATION OF DIFFERENT FORMS OF O-ACETYLSERINE SULFHYDRYLASE FROM SEEDLINGS OF 2 SPECIES OF PHASEOLUS
    BERTAGNOLLI, BL
    WEDDING, RT
    [J]. PLANT PHYSIOLOGY, 1977, 60 (01) : 115 - 121
  • [5] BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
  • [6] Complete genome sequence of the methanogenic archaeon, Methanococcus jannaschii
    Bult, CJ
    White, O
    Olsen, GJ
    Zhou, LX
    Fleischmann, RD
    Sutton, GG
    Blake, JA
    FitzGerald, LM
    Clayton, RA
    Gocayne, JD
    Kerlavage, AR
    Dougherty, BA
    Tomb, JF
    Adams, MD
    Reich, CI
    Overbeek, R
    Kirkness, EF
    Weinstock, KG
    Merrick, JM
    Glodek, A
    Scott, JL
    Geoghagen, NSM
    Weidman, JF
    Fuhrmann, JL
    Nguyen, D
    Utterback, TR
    Kelley, JM
    Peterson, JD
    Sadow, PW
    Hanna, MC
    Cotton, MD
    Roberts, KM
    Hurst, MA
    Kaine, BP
    Borodovsky, M
    Klenk, HP
    Fraser, CM
    Smith, HO
    Woese, CR
    Venter, JC
    [J]. SCIENCE, 1996, 273 (5278) : 1058 - 1073
  • [7] SULFUR METABOLISM IN PARACOCCUS-DENITRIFICANS - PURIFICATION, PROPERTIES AND REGULATION OF SERINE TRANSACETYLASE, O-ACETYLSERINE SULFYDRYLASE AND BETA-CYSTATHIONASE
    BURNELL, JN
    WHATLEY, FR
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA, 1977, 481 (01) : 246 - 265
  • [8] DNA-SEQUENCES OF THE CYSK REGIONS OF SALMONELLA-TYPHIMURIUM AND ESCHERICHIA-COLI AND LINKAGE OF THE CYSK REGIONS TO PTSH
    BYRNE, CR
    MONROE, RS
    WARD, KA
    KREDICH, NM
    [J]. JOURNAL OF BACTERIOLOGY, 1988, 170 (07) : 3150 - 3157
  • [9] CYSTEINE AND S-SULPHOCYSTEINE BIOSYNTHESIS IN BACTERIA
    CHAMBERS, LA
    TRUDINGER, PA
    [J]. ARCHIV FUR MIKROBIOLOGIE, 1971, 77 (02): : 165 - +
  • [10] COOK PF, 1976, J BIOL CHEM, V251, P2023