title: Identification of novel acetyltransferase activity on the thermostable protein ST0452 from Sulfolobus tokodaii strain 7.
abstract: A 401-residue-long protein, ST0452, has been identified from a thermophilic archaeon, Sulfolobus tokodaii strain 7, as a glucose-1-phosphate thymidylyltransferase (Glc-1-P TTase) homolog with a 170-residue-long extra C-terminus portion. Functional analyses of the ST0452 protein have confirmed that the protein possessed dual sugar-1-phosphate nucleotidylyltransferase (sugar-1-P NTase) activities. The 24 repeats of a signature motif sequence which has been found in bacterial acetyltransferases, (L/I/V)-(G/A/E/D)-XX-(S/T/A/V)-X, were detected at the C terminus of the ST0452 protein. This observation prompted our group to investigate the acetyltransferase activity of the ST0452 protein. Detection of the release of coenzyme A (CoA) from acetyl-CoA and the production of UDP-N-acetyl-d-glucosamine (UDP-GlcNAc) from glucosamine-1-phosphate (GlcN-1-P) and UTP in the presence of the ST0452 protein revealed that this protein possesses the GlcN-1-P-specific acetyltransferase activity. In addition, analyses of substrate specificity showed that acetyltransferase activity of the ST0452 protein is capable of catalyzing the change of galactosamine-1-phosphate (GalN-1-P) to N-acetyl-d-galactosamine-1-phosphate (GalNAc-1-P) as well as GlcN-1-P and that its sugar-1-P NTase activity is capable of producing UDP-GalNAc from GalNAc-1-P and UTP. This is the first report of a thermostable bifunctional enzyme with GalN-1-P acetyltransferase and GalNAc-1-P uridyltransferase activities. The observation reveals that the bacteria-type UDP-GlcNAc biosynthetic pathway from fructose-6-phospate is utilized in this archaeon and represents a novel biosynthetic pathway for producing UDP-GalNAc from GalN-1-P in this microorganism.
authors.affiliation: Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Nakoji 3-11-46, Amagasaki, Hyogo 661-0974, Japan.
authors.forename: Zilian
authors.initials: Z
authors.lastname: Zhang
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Akutsu
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Kawarabayasi
mesh_terms: D000105:Acetyl Coenzyme A / Q000378:metabolism; D000116:Acetylgalactosamine / Q000031:analogs & derivatives / Q000378:metabolism; D000123:Acetyltransferases / Q000737:chemistry / Q000145:classification / Q000378:metabolism*; D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry / Q000145:classification / Q000378:metabolism*; D003065:Coenzyme A / Q000378:metabolism; D005688:Galactosamine / Q000031:analogs & derivatives / Q000378:metabolism; D005695:Galactosephosphates / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid; D016944:Sulfolobus / Q000201:enzymology* / Q000378:metabolism; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000378:metabolism
chemical_list: D019843:Archaeal Proteins; D005695:Galactosephosphates; C115144:N-acetylgalactosamine-1-phosphate; C079528:galactosamine 1-phosphate; D014537:Uridine Diphosphate N-Acetylglucosamine; D000105:Acetyl Coenzyme A; D005688:Galactosamine; D000123:Acetyltransferases; D000116:Acetylgalactosamine; D003065:Coenzyme A
references.citation: Methods Enzymol. 1983;91:49-60
references.pmid: 6855597
references.citation: Biochemistry. 1996 Jan 16;35(2):579-85
references.pmid: 8555230
references.citation: Anal Biochem. 2008 Oct 1;381(1):94-100
references.pmid: 18634748
references.citation: Science. 1995 Nov 10;270(5238):997-1000
references.pmid: 7481807
references.citation: Biochemistry. 1997 Jan 21;36(3):489-94
references.pmid: 9012664
references.citation: Yeast. 2006 Jan 15;23(1):1-14
references.pmid: 16408321
references.citation: Microbiology (Reading). 2008 Sep;154(Pt 9):2730-2739
references.pmid: 18757806
references.citation: J Biol Chem. 1996 May 31;271(22):13147-54
references.pmid: 8662687
references.citation: Biochemistry. 2001 Feb 20;40(7):1913-21
references.pmid: 11329257
references.citation: EMBO J. 1999 Aug 2;18(15):4096-107
references.pmid: 10428949
references.citation: Biochemistry. 2006 Nov 21;45(46):13760-8
references.pmid: 17105195
references.citation: J Biol Chem. 2000 Jun 23;275(25):19060-7
references.pmid: 10747995
references.citation: J Biol Chem. 2000 Apr 28;275(17):12821-32
references.pmid: 10777580
references.citation: J Mol Biol. 2001 Jan 12;305(2):279-89
references.pmid: 11124906
references.citation: J Bacteriol. 1994 Sep;176(18):5788-95
references.pmid: 8083170
references.citation: Bioorg Med Chem Lett. 2005 Dec 15;15(24):5459-62
references.pmid: 16203137
references.citation: J Bacteriol. 1998 Sep;180(18):4799-803
references.pmid: 9733680
references.citation: J Biol Chem. 2005 Apr 8;280(14):13493-502
references.pmid: 15701638
references.citation: J Biol Chem. 2005 Mar 11;280(10):9698-705
references.pmid: 15598657
references.citation: J Biol Chem. 2000 Jan 14;275(2):1433-8
references.pmid: 10625695
references.citation: J Biol Chem. 1996 Aug 23;271(34):20776-82
references.pmid: 8702831
references.citation: J Biol Chem. 2001 Apr 13;276(15):11844-51
references.pmid: 11118459
references.citation: EMBO J. 2005 Feb 23;24(4):694-704
references.pmid: 15692569
references.citation: J Biol Chem. 1996 Jan 5;271(1):32-9
references.pmid: 8550580
references.citation: Appl Environ Microbiol. 2005 Aug;71(8):4703-12
references.pmid: 16085866
references.citation: J Biol Chem. 2005 Feb 11;280(6):4792-802
references.pmid: 15509570
references.citation: J Bacteriol. 2008 Apr;190(8):2987-96
references.pmid: 18263721
references.citation: EMBO J. 1996 May 15;15(10):2323-30
references.pmid: 8665839
references.citation: J Biol Chem. 2005 Sep 23;280(38):32784-91
references.pmid: 16006554
references.citation: Biol Reprod. 1994 May;50(5):1087-93
references.pmid: 8025165
references.citation: J Biochem. 2007 Apr;141(4):553-62
references.pmid: 17307792