Protein Engineering vol. 16 no. 11 pp. 853-860, 2003
© 2003 Oxford University Press
A broad-spectrum peptide inhibitor of ß-lactamase identified using phage display and peptide arrays
1Department of Molecular Virology and Microbiology, 2Structural and Computational Biology and Molecular Biophysics Program and 3Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA
4 To whom correspondence should be addressed. e-mail: timothyp{at}bcm.tmc.edu
Hydrolysis of ß-lactam antibiotics by ß-lactamase enzymes is the most common mechanism of bacterial resistance to these agents. Several small-molecule, mechanism-based inhibitors of ß-lactamases such as clavulanic acid are clinically available although resistance to these inhibitors has been increasing in bacterial populations. In addition, these inhibitors act only on class A ß-lactamases. Here we utilized phage display to identify peptides that bind to the class A ß-lactamase, TEM-1. The binding affinity of one of these peptides was further optimized by the synthesis of peptide arrays using SPOT synthesis technology. After two rounds of optimization, a linear 6-mer peptide with the sequence RRGHYY was obtained. A soluble version of this peptide was synthesized and found to inhibit TEM-1 ß-lactamase with a Ki of 136 µM. Surprisingly, the peptide inhibits the class A Bacillus anthracis Bla1 ß-lactamase with a Ki of 42 µM and the class C ß-lactamase, P99, with a Ki of 140 µM, despite the fact that it was not optimized to bind these enzymes. This peptide may be a useful starting point for the design of non-ß-lactam, broad-spectrum peptidomimetic inhibitors of ß-lactamases.
Received June 25, 2003; revised September 3, 2003; accepted September 9, 2003.
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