1mb9 Citations

The catalytic cycle of beta -lactam synthetase observed by x-ray crystallographic snapshots.

Proc. Natl. Acad. Sci. U.S.A. 99 14752-7 (2002)
Related entries: 1mbz, 1mc1, 1m1z

Cited: 27 times
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Abstract

The catalytic cycle of the ATP/Mg(2+)-dependent enzyme beta-lactam synthetase (beta-LS) from Streptomyces clavuligerus has been observed through a series of x-ray crystallographic snapshots. Chemistry is initiated by the ordered binding of ATP/Mg(2+) and N(2)-(carboxyethyl)-l-arginine (CEA) to the apoenzyme. The apo and ATP/Mg(2+) structures described here, along with the previously described CEA.alpha,beta-methyleneadenosine 5'-triphosphate (CEA.AMP-CPP)/Mg(2+) structure, illuminate changes in active site geometry that favor adenylation. In addition, an acyladenylate intermediate has been trapped. The substrate analog N(2)-(carboxymethyl)-l-arginine (CMA) was adenylated by ATP in the crystal and represents a close structural analog of the previously proposed CEA-adenylate intermediate. Finally, the structure of the ternary product complex deoxyguanidinoproclavaminic acid (DGPC).AMP/PP(i)/Mg(2+) has been determined. The CMA-AMP/PP(i)/Mg(2+) and DGPC.AMP/PP(i)/Mg(2+) structures reveal interactions in the active site that facilitate beta-lactam formation. All of the ATP-bound structures differ from the previously described CEA.AMP-CPP/Mg(2+) structure in that two Mg(2+) ions are found in the active sites. These Mg(2+) ions play critical roles in both the adenylation and beta-lactamization reactions.

Reviews citing this publication (8)

  1. Antibiotics from Gram-negative bacteria: a comprehensive overview and selected biosynthetic highlights. Masschelein J, Jenner M, Challis GL. Nat Prod Rep 34 712-783 (2017)
  2. Convergent biosynthetic pathways to β-lactam antibiotics. Townsend CA. Curr Opin Chem Biol 35 97-108 (2016)
  3. The enzymes of β-lactam biosynthesis. Hamed RB, Gomez-Castellanos JR, Henry L, Ducho C, McDonough MA, Schofield CJ. Nat Prod Rep 30 21-107 (2013)
  4. Origins of the β-lactam rings in natural products. Tahlan K, Jensen SE. J. Antibiot. 66 401-410 (2013)
  5. Low-molecular-weight post-translationally modified microcins. Severinov K, Semenova E, Kazakov A, Kazakov T, Gelfand MS. Mol. Microbiol. 65 1380-1394 (2007)
  6. Asparagine synthetase chemotherapy. Richards NG, Kilberg MS. Annu. Rev. Biochem. 75 629-654 (2006)
  7. Regulation and biosynthesis of carbapenem antibiotics in bacteria. Coulthurst SJ, Barnard AM, Salmond GP. Nat. Rev. Microbiol. 3 295-306 (2005)
  8. Advances in kinetic protein crystallography. Bourgeois D, Royant A. Curr. Opin. Struct. Biol. 15 538-547 (2005)

Articles citing this publication (19)

  1. Two enzymes catalyze the maturation of a lasso peptide in Escherichia coli. Duquesne S, Destoumieux-Garzón D, Zirah S, Goulard C, Peduzzi J, Rebuffat S. Chem. Biol. 14 793-803 (2007)
  2. Two sets of paralogous genes encode the enzymes involved in the early stages of clavulanic acid and clavam metabolite biosynthesis in Streptomyces clavuligerus. Tahlan K, Park HU, Wong A, Beatty PH, Jensen SE. Antimicrob. Agents Chemother. 48 930-939 (2004)
  3. iso-Migrastatin, migrastatin, and dorrigocin production in Streptomyces platensis NRRL 18993 is governed by a single biosynthetic machinery featuring an acyltransferase-less type I polyketide synthase. Lim SK, Ju J, Zazopoulos E, Jiang H, Seo JW, Chen Y, Feng Z, Rajski SR, Farnet CM, Shen B. J. Biol. Chem. 284 29746-29756 (2009)
  4. Two oligopeptide-permease-encoding genes in the clavulanic acid cluster of Streptomyces clavuligerus are essential for production of the beta-lactamase inhibitor. Lorenzana LM, Pérez-Redondo R, Santamarta I, Martín JF, Liras P. J. Bacteriol. 186 3431-3438 (2004)
  5. The enzymology of clavam and carbapenem biosynthesis. Kershaw NJ, Caines ME, Sleeman MC, Schofield CJ. Chem. Commun. (Camb.) 4251-4263 (2005)
  6. Stable analogues of OSB-AMP: potent inhibitors of MenE, the o-succinylbenzoate-CoA synthetase from bacterial menaquinone biosynthesis. Lu X, Zhou R, Sharma I, Li X, Kumar G, Swaminathan S, Tonge PJ, Tan DS. Chembiochem 13 129-136 (2012)
  7. Revisiting the steady state kinetic mechanism of glutamine-dependent asparagine synthetase from Escherichia coli. Tesson AR, Soper TS, Ciustea M, Richards NG. Arch. Biochem. Biophys. 413 23-31 (2003)
  8. A "diels-alderase" at last. Townsend CA. Chembiochem 12 2267-2269 (2011)
  9. Amidation of bioactive peptides: the structure of the lyase domain of the amidating enzyme. Chufán EE, De M, Eipper BA, Mains RE, Amzel LM. Structure 17 965-973 (2009)
  10. Dissection of the stepwise mechanism to beta-lactam formation and elucidation of a rate-determining conformational change in beta-lactam synthetase. Raber ML, Freeman MF, Townsend CA. J. Biol. Chem. 284 207-217 (2009)
  11. A critical electrostatic interaction mediates inhibitor recognition by human asparagine synthetase. Ikeuchi H, Meyer ME, Ding Y, Hiratake J, Richards NG. Bioorg. Med. Chem. 17 6641-6650 (2009)
  12. Structure of anthrax edema factor-calmodulin-adenosine 5'-(alpha,beta-methylene)-triphosphate complex reveals an alternative mode of ATP binding to the catalytic site. Shen Y, Guo Q, Zhukovskaya NL, Drum CL, Bohm A, Tang WJ. Biochem. Biophys. Res. Commun. 317 309-314 (2004)
  13. A conserved tyrosyl-glutamyl catalytic dyad in evolutionarily linked enzymes: carbapenam synthetase and beta-lactam synthetase. Raber ML, Arnett SO, Townsend CA. Biochemistry 48 4959-4971 (2009)
  14. Rate-limiting steps and role of active site Lys443 in the mechanism of carbapenam synthetase. Arnett SO, Gerratana B, Townsend CA. Biochemistry 46 9337-9345 (2007)
  15. A conserved lysine in beta-lactam synthetase assists ring cyclization: Implications for clavam and carbapenem biosynthesis. Raber ML, Castillo A, Greer A, Townsend CA. Chembiochem 10 2904-2912 (2009)
  16. A conserved glutamate controls the commitment to acyl-adenylate formation in asparagine synthetase. Meyer ME, Gutierrez JA, Raushel FM, Richards NG. Biochemistry 49 9391-9401 (2010)
  17. Substrate activation and conformational dynamics of guanosine 5'-monophosphate synthetase. Oliver JC, Linger RS, Chittur SV, Davisson VJ. Biochemistry 52 5225-5235 (2013)
  18. Engineering the synthetic potential of β-lactam synthetase and the importance of catalytic loop dynamics. Labonte JW, Kudo F, Freeman MF, Raber ML, Townsend CA. Medchemcomm 3 960-966 (2012)
  19. Exploring the role of conformational heterogeneity in cis-autoproteolytic activation of ThnT. Buller AR, Freeman MF, Schildbach JF, Townsend CA. Biochemistry 53 4273-4281 (2014)