title: A CBS domain-containing pyrophosphatase of Moorella thermoacetica is regulated by adenine nucleotides.
abstract: CBS (cystathionine beta-synthase) domains are found in proteins from all kingdoms of life, and point mutations in these domains are responsible for a variety of hereditary diseases in humans; however, the functions of CBS domains are not well understood. In the present study, we cloned, expressed in Escherichia coli, and characterized a family II PPase (inorganic pyrophosphatase) from Moorella thermoacetica (mtCBS-PPase) that has a pair of tandem 60-amino-acid CBS domains within its N-terminal domain. Because mtCBS-PPase is a dimer and requires transition metal ions (Co2+ or Mn2+) for activity, it resembles common family II PPases, which lack CBS domains. The mtCBS-PPase, however, has lower activity than common family II PPases, is potently inhibited by ADP and AMP, and is activated up to 1.6-fold by ATP. Inhibition by AMP is competitive, whereas inhibition by ADP and activation by ATP are both of mixed types. The nucleotides are effective at nanomolar (ADP) or micromolar concentrations (AMP and ATP) and appear to compete for the same site on the enzyme. The nucleotide-binding affinities are thus 100-10000-fold higher than for other CBS-domain-containing proteins. Interestingly, genes encoding CBS-PPase occur most frequently in bacteria that have a membrane-bound H+-translocating PPase with a comparable PP(i)-hydrolysing activity. Our results suggest that soluble nucleotide-regulated PPases act as amplifiers of metabolism in bacteria by enhancing or suppressing ATP production and biosynthetic reactions at high and low [ATP]/([AMP]+[ADP]) ratios respectively.
authors: Department of Biochemistry and Food Chemistry, University of Turku, FIN-20014 Turku, Finland.
authors: Joonas
authors: J
authors: Jämsen
authors: Heidi
authors: H
authors: Tuominen
authors: Anu
authors: A
authors: Salminen
authors: Georgiy A
authors: GA
authors: Belogurov
authors: Natalia N
authors: NN
authors: Magretova
authors: Alexander A
authors: AA
authors: Baykov
authors: Reijo
authors: R
authors: Lahti
mesh_terms: D000227:Adenine Nucleotides / Q000378:metabolism*; D002384:Catalysis; D003001:Cloning, Molecular; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D043564:Inorganic Pyrophosphatase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D013379:Substrate Specificity; D046950:Thermoanaerobacterium / Q000201:enzymology*
chemical_list: D000227:Adenine Nucleotides; D043564:Inorganic Pyrophosphatase
references: J Bacteriol. 1990 Oct;172(10):5686-9
references: 2170325
references: J Bacteriol. 1984 Aug;159(2):700-3
references: 6746575
references: Trends Biochem Sci. 1992 Jul;17(7):262-6
references: 1323891
references: Plant J. 1992 Jul;2(4):571-81
references: 1344891
references: Res Microbiol. 2004 Dec;155(10):869-83
references: 15630808
references: J Biol Chem. 2005 Sep 16;280(37):32452-8
references: 16027167
references: Am J Physiol Cell Physiol. 2005 Dec;289(6):C1369-78
references: 16275737
references: Structure. 2006 Feb;14(2):299-307
references: 16472749
references: Proc Natl Acad Sci U S A. 2006 Jul 11;103(28):10624-9
references: 16815971
references: Biochem J. 2006 Sep 1;398(2):289-94
references: 16686597
references: Nat Struct Mol Biol. 2007 Jan;14(1):60-7
references: 17195847
references: J Biol Chem. 1995 Dec 22;270(51):30804-12
references: 8530523
references: Trends Biochem Sci. 1997 Jan;22(1):12-3
references: 9020585
references: Trends Biochem Sci. 1998 Jan;23(1):17-9
references: 9478130
references: Arch Biochem Biophys. 1998 Jul 15;355(2):222-32
references: 9675031
references: Microbiology. 1998 Sep;144 ( Pt 9):2563-71
references: 9782505
references: FEBS Lett. 1998 Nov 20;439(3):263-6
references: 9845334
references: Biochemistry. 1999 Apr 13;38(15):4691-700
references: 10200156
references: Prog Mol Subcell Biol. 1999;23:127-50
references: 10448675
references: Res Microbiol. 2000 Mar;151(2):129-34
references: 10865958
references: Arch Biochem Biophys. 2000 Jul 15;379(2):292-8
references: 10898947
references: Biochemistry. 2000 Aug 29;39(34):10548-56
references: 10956046
references: J Biol Chem. 2001 Jul 6;276(27):24511-8
references: 11342544
references: Structure. 2001 Apr 4;9(4):289-97
references: 11525166
references: J Mol Biol. 2001 Nov 2;313(4):797-811
references: 11697905
references: Biochem J. 2002 Nov 1;367(Pt 3):901-6
references: 12169093
references: Biochem Biophys Res Commun. 2003 Oct 17;310(2):600-5
references: 14521953
references: Nucleic Acids Res. 2004 Jan 1;32(Database issue):D138-41
references: 14681378
references: J Clin Invest. 2004 Jan;113(2):182-4
references: 14722609
references: J Clin Invest. 2004 Jan;113(2):274-84
references: 14722619
references: Biochemistry. 2004 Feb 3;43(4):1065-74
references: 14744152
references: Proteins. 2004 Oct 1;57(1):213-7
references: 15326606
references: Physiol Genomics. 2004 Sep 16;19(1):74-83
references: 15252188
references: Chem Rev. 1966 Oct;66(5):501-27
references: 4288270
references: Anal Biochem. 1981 Sep 1;116(1):1-4
references: 6118076
references: J Biol Chem. 1991 Jul 5;266(19):12168-72
references: 1648084