This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Majiduddin, F. K.
Right arrow Articles by Palzkill, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Majiduddin, F. K.
Right arrow Articles by Palzkill, T.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, March 2003, p. 1062-1067, Vol. 47, No. 3
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.3.1062-1067.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Amino Acid Sequence Requirements at Residues 69 and 238 for the SME-1 ß-Lactamase To Confer Resistance to ß-Lactam Antibiotics

Fahd K. Majiduddin1 and Timothy Palzkill1,2*

Verna and Marrs McLean Department of Biochemistry and Molecular Biology,1 Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas 770302

Received 4 September 2002/ Returned for modification 24 October 2002/ Accepted 6 December 2002

Carbapenem antibiotics have been used to counteract resistant strains of bacteria harboring ß-lactamases and extended-spectrum ß-lactamases. Four enzymes from the class A group of ß-lactamases, NMC-A, IMI-1, SME-1, and KPC-1, efficiently hydrolyze carbapenem antibiotics. Sequence comparisons and structural information indicate that cysteines at amino acid residues 69 and 238, which are conserved in all four of these enzymes, form a disulfide bond that is unique to these ß-lactamases. To test whether this disulfide bond is required for catalytic activity, the codons for residues Cys69 and Cys238 were randomized individually and simultaneously by PCR-based mutagenesis to create random replacement libraries for these positions. Mutants that were able to confer resistance to ampicillin, imipenem, or cefotaxime were selected from these libraries. The results indicate that positions Cys69 and Cys238 are critical for hydrolysis of all of the antibiotics tested, suggesting that the disulfide bond is generally required for this enzyme to catalyze the hydrolysis of ß-lactam antibiotics.

* Corresponding author. Mailing address: Department of Molecular Virology and Microbiology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030. Phone: (713) 798-5609. Fax: (713) 798-7375. E-mail: timothyp{at}bcm.tmc.edu.

Antimicrobial Agents and Chemotherapy, March 2003, p. 1062-1067, Vol. 47, No. 3
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.3.1062-1067.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Walther-Rasmussen, J., Hoiby, N. (2007). Class A carbapenemases. J Antimicrob Chemother 60: 470-482 [Abstract] [Full Text]  
  • Marciano, D. C., Karkouti, O. Y., Palzkill, T. (2007). A Fitness Cost Associated With the Antibiotic Resistance Enzyme SME-1 {beta}-Lactamase. Genetics 176: 2381-2392 [Abstract] [Full Text]  
  • Queenan, A. M., Bush, K. (2007). Carbapenemases: the Versatile {beta}-Lactamases. Clin. Microbiol. Rev. 20: 440-458 [Abstract] [Full Text]  
  • Majiduddin, F. K., Palzkill, T. (2005). Amino Acid Residues That Contribute to Substrate Specificity of Class A {beta}-Lactamase SME-1. Antimicrob. Agents Chemother. 49: 3421-3427 [Abstract] [Full Text]  
  • Weiss, W. J., Petersen, P. J., Murphy, T. M., Tardio, L., Yang, Y., Bradford, P. A., Venkatesan, A. M., Abe, T., Isoda, T., Mihira, A., Ushirogochi, H., Takasake, T., Projan, S., O'Connell, J., Mansour, T. S. (2004). In Vitro and In Vivo Activities of Novel 6-Methylidene Penems as {beta}-Lactamase Inhibitors. Antimicrob. Agents Chemother. 48: 4589-4596 [Abstract] [Full Text]  
  • Zhang, Z., Palzkill, T. (2003). Determinants of Binding Affinity and Specificity for the Interaction of TEM-1 and SME-1 {beta}-Lactamase with {beta}-Lactamase Inhibitory Protein. J. Biol. Chem. 278: 45706-45712 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»