[en] beta-Lactamases are the main cause of bacterial resistance to penicillins and cephalosporins. Class A beta-lactamases, the largest group of beta-lactamases, have been found in many bacterial strains, including mycobacteria, for which no beta-lactamase structure has been previously reported. The crystal structure of the class A beta-lactamase from Mycobacterium fortuitum (MFO) has been solved at 2.13-A resolution. The enzyme is a chromosomally encoded broad-spectrum beta-lactamase with low specific activity on cefotaxime. Specific features of the active site of the class A beta-lactamase from M. fortuitum are consistent with its specificity profile. Arg278 and Ser237 favor cephalosporinase activity and could explain its broad substrate activity. The MFO active site presents similarities with the CTX-M type extended-spectrum beta-lactamases but lacks a specific feature of these enzymes, the VNYN motif (residues 103 to 106), which confers on CTX-M-type extended-spectrum beta-lactamases a more efficient cefotaximase activity.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Sauvage, Eric ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Fonze, Eveline
Quinting, Birgit
Galleni, Moreno ; Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques
Frère, Jean-Marie ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Charlier, Paulette ; Université de Liège - ULiège > Département des sciences de la vie > Cristallographie des macromolécules biologiques
Language :
English
Title :
Crystal structure of the Mycobacterium fortuitum class A beta-lactamase: structural basis for broad substrate specificity.
Publication date :
2006
Journal title :
Antimicrobial Agents and Chemotherapy
ISSN :
0066-4804
eISSN :
1098-6596
Publisher :
American Society for Microbiology (ASM), Washington, United States - District of Columbia
Amicosante, G., N. Franceschini, B. Segatore, A. Oratore, L. Fattorini, G. Orefici, J. Van Beeumen, and J. M. Frere. 1990. Characterization of a beta-lactamase produced in Mycobacterium fortuitum D316. Biochcm. J. 271:729-734.
Bonnet, R. 2004. Growing group of extended-spectrum beta-lactamases: the CTX-M enzymes. Antimicrob. Agents Chemother. 48:1-14.
Cantu, C., III, W. Huang, and T. Palzkill. 1997. Cephalosporin substrate specificity determinants of TEM-1 beta-lactamase. J. Biol. Chem. 272:29144-29150.
CCP4. 1994. The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D 50:760-763.
Chen, Y., J. Delmas, J. Sirot, B. Shoichet, and R. Bonnet. 2005. Atomic resolution structures of CTX-M beta-lactamases: extended spectrum activities from increased mobility and decreased stability. J. Mol. Biol. 348:349-362.
Dideberg, O., P. Charlier, J. P. Wety, P. Dehottay, J. Dusart, T. Erpicum, J. M. Frere, and J. M. Ghuysen. 1987. The crystal structure of the beta-lactamase of Streptomyces albus G at 0.3 nm resolution. Biochem. J. 245:911-913.
Dive, G., and D. Dehareng. 1999. Serine peptidase catalytic machinery: cooperative one-step mechanism. Int. J. Quant. Chem. 73:161-174.
Emsley, P., and K. Cowtan. 2004. Coot: model-building tools for molecular graphics. Acta Crystallogr. D 60:2126-2132.
Fattorini, L., G. Orefici, S. H. Jin, G. Scardaci, G. Amicosante, N. Franceschini, and I. Chopra. 1992. Resistance to beta-lactams in Mycobacterium fortuitum. Antimicrob. Agents Chemother. 36:1068-1072.
Fonze, E., M. Vanhove, G. Dive, E. Sauvage, J. M. Frere, and P. Charlier. 2002. Crystal structures of the Bacillus licheniformis BS3 class A beta-lactamase and of the acyl-enzyme adduct formed with cefoxitin. Biochemistry 41:1877-1885.
Frere, J. M., C. Dormans, V. M. Lenzini, and C. Duyckaerts. 1982. Interaction of clavulanate with the beta-lactamases of Streptomyces albus G and Actinomadura R39. Biochem. J. 207:429-436.
Galleni, M., N. Franceschini, B. Quinting, L. Fattorini, G. Orefici, A. Oratore, J. M. Frere, and G. Amicosante. 1994. Use of the chromosomal class A beta-lactamase of Mycobacterium fortuitum D316 to study potentially poor substrates and inhibitory beta-lactam compounds. Antimicrob. Agents Chemother. 38:1608-1614.
Gazouli, M., N. J. Legakis, and L. S. Tzouvelekis. 1998. Effect of substitution of Asn for Arg-276 in the cefotaxime-hydrolyzing class A beta-lactamase CTX-M-4. FEMS Microbiol. Lett. 169:289-293.
Guillaume, G., M. Vanhove, J. Lamotte-Brasseur, P. Ledent, M. Jamin, B. Joris, and J. M. Frere. 1997. Site-directed mutagenesis of glutamate 166 in two beta-lactamases. Kinetic and molecular modeling studies. J. Biol. Chem. 272:5438-5444.
Hackbarth, C. J., I. Unsal, and H. F. Chambers. 1997. Cloning and sequence analysis of a class A beta-lactamase from Mycobacterium tuberculosis H37Ra. Antimicrob. Agents Chemother. 41:1182-1185.
Healey, W. J., M. R. Labgold, and J. H. Richards. 1989. Substrate specificities in class A beta-lactamases: preference for penams vs. cephems. The role of residue 237. Proteins 6:275-283.
Hokenson, M. J., G. A. Cope, E. R. Lewis, K. A. Oberg, and A. L. Fink. 2000. Enzyme-induced strain/distortion in the ground-state ES complex in beta-lactamase catalysis revealed by FTIR. Biochemistry 39:6538-6545.
Ibuka, A. S., Y. Ishii, M. Galleni, M. Ishiguro, K. Yamaguchi, J. M. Frere, H. Matsuzawa, and H. Sakai. 2003. Crystal structure of extended-spectrum beta-lactamase Toho-1: insights into the molecular mechanism for catalytic reaction and substrate specificity expansion. Biochemistry 42:10634-10643.
Imtiaz, U., E. K. Manavathu, S. Mobashery, and S. A. Lerner. 1994. Reversal of clavulanate resistance conferred by a Ser-244 mutant of TEM-1 beta-lactamase as a result of a second mutation (Arg to Ser at position 164) that enhances activity against ceftazidime. Antimicrob. Agents Chemother. 38:1134-1139.
Jacob, F., B. Joris, S. Lepage, J. Dusart, and J. M. Frere. 1990. Role of the conserved amino acids of the SDN′ loop (Ser130, Asp131 and Asn132) in a class A beta-lactamase studied by site-directed mutagenesis. Biochem. J. 271:399-406.
Jarlier, V., and H. Nikaido. 1994. Mycobacterial cell wall: structure and role in natural resistance to antibiotics. FEMS Microbiol. Lett. 123:11-18.
Laskowski, R. A. 2001. PDBsum: summaries and analyses of PDB structures. Nucleic Acids Res. 29:221-222.
Leslie, A. G. W. 1991. Molecular data processing. Crystallogr. Comput. 5:50-61.
Liu, J., C. E. Barry III, G. S. Besra, and H. Nikaido. 1996. Mycolic acid structure determines the fluidity of the mycobacterial cell wall. J. Biol. Chem. 271:29545-29551.
Maveyraud, L., I. Saves, O. Burlet-Schiltz, P. Swaren, J. M. Masson, M. Delaire, L. Mourey, J. C. Prome, and J. P. Samama. 1996. Structural basis of extended spectrum TEM beta-lactamases. Crystallographic, kinetic, and mass spectrometric investigations of enzyme mutants. J. Biol. Chem. 271:10482-10489.
Navaza, J. 2001. Implementation of molecular replacement in AMoRc. Acta Crystallogr. D 57:1367-1372.
Nukaga, M., K. Mayama, G. V. Crichlow, and J. R. Knox. 2002. Structure of an extended-spectrum class A beta-lactamase from Proteus vulgaris K1. J. Mol. Biol. 317:109-117.
Oliva, M., O. Dideberg, and M. J. Field. 2003. Understanding the acylation mechanisms of active-site serine penicillin-recognizing proteins: a molecular dynamics simulation study. Proteins 53:88-100.
Orencia, M. C., J. S. Yoon, J. E. Ness, W. P. Stemmer, and R. C. Stevens. 2001. Predicting the emergence of antibiotic resistance by directed evolution and structural analysis. Nat. Struct. Biol. 8:238-242.
Perez-Llarena, F., J. F. Martin, M. Galleni, J. J. Coque, J. L. Fuente, J. M. Frere, and P. Liras. 1997. The bla gene of the cephamycin cluster of Streptomyces clavuligerus encodes a class A beta-lactamase of low enzymatic activity. J. Bacteriol. 179:6035-6040.
Petrella, S., D. Clermont, I. Casin, V. Jarlier, and W. Sougakoff. 2001. Novel class A beta-lactamase Sed-1 from Citrobacter sedlakii: genetic diversity of beta-lactamases within the Citrobacter genus. Antimicrob. Agents Chemother. 45:2287-2298.
Petrosino, J. F., M. Baker, and T. Palzkill. 1999. Susceptibility of beta-lactamase to core amino acid substitutions. Protein Eng. 12:761-769.
Poirel, L., F. Laurent, T. Naas, R. Labia, P. Boiron, and P. Nordmann. 2001. Molecular and biochemical analysis of AST-1, a class A beta-lactamase from Nocardia asteroides sensu stricto. Antimicrob. Agents Chemother. 45:878-882.
Poirel, L., T. Naas, I. Le Thomas, A. Karim, E. Bingen, and P. Nordmann. 2001. CTX-M-type extended-spectrum beta-lactamase that hydrolyzes ceftazidime through a single amino acid substitution in the omega loop. Antimicrob. Agents Chemother. 45:3355-3361.
Quinting, B., J. M. Reyrat, D. Monnaie, G. Amicosante, V. Pelicic, B. Gicquel, J. M. Frere, and M. Galleni. 1997. Contribution of beta-lactamase production to the resistance of mycobacteria to beta-lactam antibiotics. FEBS Lett. 406:275-278.
Raquet, X., J. Lamotte-Brasseur, E. Fonze, S. Goussard, P. Courvalin, and J. M. Frere. 1994. TEM beta-lactamase mutants hydrolysing third-generation cephalosporins. A kinetic and molecular modelling analysis. J. Mol. Biol. 244:625-639.
Seoane, A., and J. M. Garcia Lobo. 1991. Cloning of chromosomal beta-lactamase genes from Yersinia enterocolitica. J. Gen. Microbiol. 137:141-146.
Shimamura, T., A. Ibuka, S. Fushinobu, T. Wakagi, M. Ishiguro, Y. Ishii, and H. Matsuzawa. 2002. Acyl-intermediate structures of the extended-spectrum class A beta-lactamase, Toho-1, in complex with cefotaxime, cephalothin, and benzylpenicillin. J. Biol. Chem. 277:46601-46608.
Sougakoff, W., T. Naas, P. Nordmann, E. Collatz, and V. Jarlier. 1999. Role of ser-237 in the substrate specificity of the carbapenem-hydrolyzing class A beta-lactamase Sme-1. Biochim. Biophys. Acta 1433:153-158.
Tamaki, M., M. Nukaga, and T. Sawai. 1994. Replacement of serine 237 in class A beta-lactamase of Proteus vulgaris modifies its unique substrate specificity. Biochemistry 33:10200-10206.
Thami, G. P., S. Kaur, J. Chander, and A. K. Attri. 2002. Post surgical atypical mycobacterial infection due to Mycobacterium fortuitum. J. Infect. 45:210-211.
Timm, J., M. G. Perilli, C. Duez, J. Trias, G. Orefici, L. Fattorini, G. Amicosante, A. Oratore, B. Joris, J. M. Frere, et al. 1994. Transcription and expression analysis, using lacZ and phoA gene fusions, of Mycobacterium fortuitum beta-lactamase genes cloned from a natural isolate and a high-level beta-lactamase producer. Mol. Microbiol. 12:491-504.
Voladri, R. K., D. L. Lakey, S. H. Hennigan, B. E. Menzies, K. M. Edwards, and D. S. Kernodle. 1998. Recombinant expression and characterization of the major beta-lactamase of Mycobacterium tuberculosis. Antimicrob. Agents Chemother. 42:1375-1381.
Wallace, R. J., Jr., J. M. Swenson, V. A. Silcox, R. C. Good, J. A. Tschen, and M. S. Stone. 1983. Spectrum of disease due to rapidly growing mycobacteria. Rev. Infect. Dis. 5:657-679.
Zafaralla, G., E. K. Manavathu, S. A. Lerner, and S. Mobashery. 1992. Elucidation of the role of arginine-244 in the turnover processes of class A beta-lactamases. Biochemistry 31:3847-3852.