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Antimicrobial Agents and Chemotherapy, November 1998, p. 2893-2897, Vol. 42, No. 11
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Display of Functional beta -Lactamase Inhibitory Protein on the Surface of M13 Bacteriophage

Wanzhi Huang,1 Joseph Petrosino,1 and Timothy Palzkill1,2,*

Department of Microbiology and Immunology1 and Department of Biochemistry,2 Baylor College of Medicine, Houston, Texas 77030

Received 3 April 1998/Returned for modification 23 July 1998/Accepted 10 August 1998

The display of proteins on the surface of filamentous phage has been shown to be a powerful method to select variants of a protein with altered binding properties from large combinatorial libraries of mutants. The beta -lactamase inhibitory protein (BLIP) is a 165-amino-acid protein that binds and inhibits TEM-1 beta -lactamase-catalyzed hydrolysis of the penicillin and cephalosporin antibiotics. Here we describe the construction of a new phagemid vector and the use of this vector to display BLIP on the surface of filamentous phage. It is shown that BLIP-displaying phage bind to immobilized beta -lactamase and that the binding can be competed off by the addition of soluble beta -lactamase. In addition, a two-step phage enzyme-linked immunosorbent assay procedure was used to demonstrate that the BLIP-displaying phage bind beta -lactamase with a 50% inhibitory concentration of 1 nM, which compares favorably with a previously published Ki of 0.6 nM. A system has therefore been established for protein engineering of BLIP to expand its range of binding to other beta -lactamases and penicillin-binding proteins.

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

Antimicrobial Agents and Chemotherapy, November 1998, p. 2893-2897, Vol. 42, No. 11
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Zhang, Z., Palzkill, T. (2004). Dissecting the Protein-Protein Interface between {beta}-Lactamase Inhibitory Protein and Class A {beta}-Lactamases. J. Biol. Chem. 279: 42860-42866 [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]  
  • Huang, W., Beharry, Z., Zhang, Z., Palzkill, T. (2003). A broad-spectrum peptide inhibitor of {beta}-lactamase identified using phage display and peptide arrays. Protein Eng Des Sel 16: 853-860 [Abstract] [Full Text]  
  • Rudgers, G. W., Palzkill, T. (1999). Identification of Residues in beta -Lactamase Critical for Binding beta -Lactamase Inhibitory Protein. J. Biol. Chem. 274: 6963-6971 [Abstract] [Full Text]  
  • Petrosino, J., Rudgers, G., Gilbert, H., Palzkill, T. (1999). Contributions of Aspartate 49 and Phenylalanine 142 Residues of a Tight Binding Inhibitory Protein of beta -Lactamases. J. Biol. Chem. 274: 2394-2400 [Abstract] [Full Text]  
  • Huang, W., Zhang, Z., Palzkill, T. (2000). Design of Potent beta -Lactamase Inhibitors by Phage Display of beta -Lactamase Inhibitory Protein. J. Biol. Chem. 275: 14964-14968 [Abstract] [Full Text]  


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