[en] The role of the mobile loop comprising residues 60-66 in metallo-beta-lactamases has been studied by site-directed mutagenesis, determination of kinetic parameters for six substrates and two inhibitors, pre-steady-state characterization of the interaction with chromogenic nitrocefin, and molecular modeling. The W64A mutation was performed in IMP-1 and BcII (after replacement of the Bcll 60-66 peptide by that of IMP-1) and always resulted in increased K-i and K. and decreased k(cat)/K-m values, an effect reinforced by complete deletion of the loop. k(cat) values were, by contrast, much more diversely affected, indicating that the loop does not systematically favor the best relative positioning of substrate and enzyme catalytic groups. The hydrophobic nature of the ligand is also crucial to strong interactions with the loop, since imipenem was almost insensitive to loop modifications.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Moali, Catherine; Université de Liège - ULiège > Centres généraux > Centre d'ingénierie des protéines
Anne, Christine ; Université de Liège - ULiège > Centres généraux > Centre d'ingénierie des protéines
Lamotte-Brasseur, Josette ; Université de Liège - ULiège > Centres généraux > Centre d'ingénierie des protéines
Groslambert, Sylvie ; Université de Liège - ULiège > Department of Chemical Engineering > PEPs (Product, Environment, Processes)
Devreese, B.
Van Beeumen, J.
Galleni, Moreno ; Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques
Frère, Jean-Marie ; Université de Liège - ULiège > Département des sciences de la vie > Département des sciences de la vie
Language :
English
Title :
Analysis of the importance of the metallo-beta-lactamase active site loop in substrate binding and catalysis
Kuwabara S., Abraham E.P. Some properties of two extracellular beta-lactamases from Bacillus cereus 569/H. Biochem. J. 103:1967;27C-30C.
Fabiane S.M., Sohi M.K., Wan T., Payne D.J., Bateson J.H., Mitchell T., Sutton B.J. Crystal structure of the zinc-dependent beta-lactamase from Bacillus cereus at 1.9 A resolution. binuclear active site with features of a mononuclear enzyme Biochemistry. 37:1998;12404-12411.
Orellano E.G., Girardini J.E., Cricco J.A., Ceccarelli E.A., Vila A.J. Spectroscopic characterization of a binuclear metal site in Bacillus cereus beta-lactamase II. Biochemistry. 37:1998;10173-10180.
Paul-Soto R., Bauer R., Frère J.M., Galleni M., Meyer-Klaucke W., Nolting H., Rossolini G.M., de Seny D., Hernandez-Valladares M., Zeppezauer M.et al. Mono- and binuclear Zn2+-beta-lactamase. Role of the conserved cysteine in the catalytic mechanism. J. Biol. Chem. 274:1999;13242-13249.
Concha N.O., Rasmussen B.A., Bush K., Herzberg O. Crystal structure of the wide-spectrum binuclear zinc beta-lactamase from Bacteroides fragilis. Structure. 4:1996;823-836.
Paul-Soto R., Hernadez-Valladares M., Galleni M., Bauer R., Zeppezauer M., Frère J.M., Adolph H.W. Mono- and binuclear Zn-beta-lactamase from Bacteroides fragilis. catalytic and structural roles of the zinc ions FEBS Lett. 438:1998;137-140.
Yang Y., Keeney D., Tang X., Canfield N., Rasmussen B.A. Kinetic properties and metal content of the metallo-beta-lactamase CcrA harboring selective amino acid substitutions. J. Biol. Chem. 274:1999;15706-15711.
Wang Z., Fast W., Benkovic S.J. On the mechanism of the metallo-beta-lactamase from Bacteroides fragilis. Biochemistry. 38:1999;10013-10023.
Laraki N., Franceschini N., Rossolini G.M., Santucci P., Meunier C., de Pauw E., Amicosante G., Frère J.M., Galleni M. Biochemical characterization of the Pseudomonas aeruginosa 101/1477 metallo-beta-lactamase IMP-1 produced by Escherichia coli. Antimicrob. Agents Chemother. 43:1999;902-906.
Concha N.O., Janson C.A., Rowling P., Pearson S., Cheever C.A., Clarke B.P., Lewis C., Galleni M., Frère J.M., Payne D.J.et al. Crystal structure of the IMP-1 metallo beta-lactamase from Pseudomonas aeruginosa and its complex with a mercaptocarboxylate inhibitor. binding determinants of a potent, broad-spectrum inhibitor Biochemistry. 39:2000;4288-4298.
Haruta S., Yamaguchi H., Yamamoto E.T., Eriguchi Y., Nukaga M., O'Hara K., Sawai T. Functional analysis of the active site of a metallo-beta-lactamase proliferating in Japan. Antimicrob. Agents Chemother. 44:2000;2304-2309.
Hernandez Valladares M., Felici A., Weber G., Adolph H.W., Zeppezauer M., Rossolini G.M., Amicosante G., Frère J.M., Galleni M. Zn(II) dependence of the Aeromonas hydrophila AE036 metallo-beta-lactamase activity and stability. Biochemistry. 36:1997;11534-11541.
Crowder M.W., Walsh T.R., Banovic L., Pettit M., Spencer J. Overexpression, purification, and characterization of the cloned metallo-beta-lactamase L1 from Stenotrophomonas maltophilia. Antimicrob. Agents Chemother. 42:1998;921-926.
Ullah J.H., Walsh T.R., Taylor I.A., Emery D.C., Verma C.S., Gamblin S.J., Spencer J. The crystal structure of the L1 metallo-beta-lactamase from Stenotrophomonas maltophilia at 1.7 A resolution. J. Mol. Biol. 284:1998;125-136.
Mercuri P.S., Bouillenne F., Boschi L., Lamotte-Brasseur J., Amicosante G., Devreese B., van Beeumen J., Frère J.M., Rossolini G.M., Galleni M. Biochemical characterization of the FEZ-1 metallo-beta-lactamase of Legionella gormanii ATCC 33297(T) produced in Escherichia coli. Antimicrob. Agents Chemother. 45:2001;1254-1262.
Carfi A., Duee E., Paul-Soto R., Galleni M., Frère J.M., Dideberg O. X-ray structure of the ZnII beta-lactamase from Bacteroides fragilis in an orthorhombic crystal form. Acta Crystallogr. D Biol. Crystallogr. 54:1998;45-57.
Carfi A., Duee E., Galleni M., Frère J.M., Dideberg O. 1.85 Å resolution structure of the zinc (II) beta-lactamase from Bacillus cereus. Acta Crystallogr. D Biol. Crystallogr. 54:1998;313-323.
Fitzgerald P.M., Wu J.K., Toney J.H. Unanticipated inhibition of the metallo-beta-lactamase from Bacteroides fragilis by 4-morpholineethanesulfonic acid (MES). a crystallographic study at 1.85-Å resolution Biochemistry. 37:1998;6791-6800.
Carfi A., Pares S., Duee E., Galleni M., Duez C., Frère J.M., Dideberg O. The 3-D structure of a zinc metallo-beta-lactamase from Bacillus cereus reveals a new type of protein fold. EMBO J. 14:1995;4914-4921.
Wang Z., Fast W., Valentine A.M., Benkovic S.J. Metallo-beta-lactamase. structure and mechanism Curr. Opin. Chem. Biol. 3:1999;614-622.
Wang Z., Fast W., Benkovic S.J. Direct observation of an enzyme bound intermediate in the catalytic cycle of the metallo-beta-lactamase from Bacteroides fragilis. J. Am. Chem. Soc. 120:1998;10788-10789.
McManus-Munoz S., Crowder M.W. Kinetic mechanism of metallo-beta-lactamase L1 from Stenotrophomonas maltophilia. Biochemistry. 38:1999;1547-1553.
Toney J.H., Fitzgerald P.M., Grover-Sharma N., Olson S.H., May W.J., Sundelof J.G., Vanderwall D.E., Cleary K.A., Grant S.K., Wu J.K.et al. Antibiotic sensitization using biphenyl tetrazoles as potent inhibitors of Bacteroides fragilis metallo-beta-lactamase. Chem. Biol. 5:1998;185-196.
Prosperi-Meys C., Wouters J., Galleni M., Lamotte-Brasseur J. Substrate binding and catalytic mechanism of class B beta-lactamases. a molecular modelling study Cell. Mol. Life Sci. 58:2001;2136-2143.
Galleni M., Lamotte-Brasseur J., Rossolini G.M., Spencer J., Dideberg O., Frère J.M. Standard numbering scheme for class B beta-lactamases. Antimicrob. Agents Chemother. 45:2001;660-663.
Scrofani S.D., Chung J., Huntley J.J., Benkovic S.J., Wright P.E., Dyson H.J. NMR characterization of the metallo-beta-lactamase from bacteroides fragilis and its interaction with a tight-binding inhibitor. role of an active-site loop Biochemistry. 38:1999;14507-14514.
Salsbury F.R. Jr., Crowley M.F., Brooks C.L. 3rd. Modeling of the metallo-beta-lactamase from B. fragilis. structural and dynamic effects of inhibitor binding Proteins. 44:2001;448-459.
Huntley J.J., Scrofani S.D., Osborne M.J., Wright P.E., Dyson H.J. Dynamics of the metallo-beta-lactamase from Bacteroides fragilis in the presence and absence of a tight-binding inhibitor. Biochemistry. 39:2000;13356-13364.
Mollard C., Moali C., Papamicael C., Damblon C., Vessilier S., Amicosante G., Schofield C.J., Galleni M., Frère J.M., Roberts G.C. Thiomandelic acid, a broad spectrum inhibitor of zinc beta-lactamases. kinetic and spectroscopic studies J. Biol. Chem. 276:2001;45015-45023.
Bicknell R., Waley S.G. Cryoenzymology of Bacillus cereus beta-lactamase II. Biochemistry. 24:1985;6876-6887.
Matagne A., Ledent P., Monnaie D., Felici A., Jamin M., Raquet X., Galleni M., Klein D., François I., Frère J.-M. Kinetic study of interaction between BRL 42715, β-lactamases, and D-Alanyl-D-Alanine peptidases. Antimicrob. Agents Chemother. 39:1995;227-231.
Carenbauer A.L., Garrity J.D., Periyannan G., Yates R.B., Crowder M.W. Probing substrate binding to metallo-β-lactamase L1 from Stenotrophomonas maltophilia by using site-directed mutagenesis. BMC Biochem. 3:2002;4.
Fast W., Wang Z., Benkovic S.J. Familial mutations and zinc stoichiometry determine the rate-limiting step of nitrocefin hydrolysis by metallo-beta-lactamase from Bacteroides fragilis. Biochemistry. 40:2001;1640-1650.
Docquier J.D., Lamotte-Brasseur J., Galleni M., Amicosante G., Frère J.-M., Rossolini G.M. On functional and structural heterogeneity of VIM-type metallo-β-lactamases. J. Antimicrob. Chemother. 51:2003;257-266.
de Seny D., Prosperi-Meys C., Bebrone C., Rossolini G.M., Page M.I., Noel P., Frère J.M., Galleni M. Mutational analysis of the two zinc binding sites of the Bacillus cereus 569/H/9 metallo-beta-lactamase. Biochem. J. 363:2002;687-696.
Haruta S., Yamamoto E., Eriguchi Y., Sawai T. Characterization of the active-site residues asparagine 167 and lysine 161 in the IMP-1 metallo beta-lacatamase. FEMS Microbiol. Lett. 197:2001;85-89.
De Meester F., Joris B., Reckinger G., Bellefroid-Bourgignon C., Frère J.M. Automated analysis of enzyme inactivation phenomena. Application to beta-lactamases and DD-peptidases Biochem. Pharmacol. 36:1987;2393-2403.