Previous Article | Next Article ![]()
Antimicrobial Agents and Chemotherapy, May 1999, p. 1124-1128, Vol. 43, No. 5
Lilly Research Laboratories, Eli Lilly and
Company, Indianapolis, Indiana 46285-0438,1
and Molecular Probes, Inc., Eugene, Oregon
974022
Received 30 July 1998/Returned for modification 5 November
1998/Accepted 2 March 1999
We describe a new, sensitive, rapid, and nonradioactive method
involving the use of the commercially available BOCILLIN FL, a
fluorescent penicillin, as a labeling reagent for the detection and
study of penicillin-binding proteins (PBPs). This method allowed rapid
detection of 30 ng of a purified PBP protein under UV light and of 2 to
4 ng of the protein with the aid of a FluorImager. This method also
allowed rapid determination of the PBP profiles of Escherichia
coli, Pseudomonas aeruginosa, and Streptococcus pneumoniae. The PBP profiles obtained are virtually
identical to those reported previously with 3H-,
14C-, or 125I-labeled penicillin. Using this
method enabled us to determine the 50% inhibitory concentrations of
the penicillin-sensitive and -resistant PBP2x proteins of S. pneumoniae for penicillin G, thereby allowing a direct evaluation
of their relative affinities for penicillin G. Finally, this method
also allowed us to compare relative affinities of a PBP2x protein
for different
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
BOCILLIN FL, a Sensitive and
Commercially Available Reagent for Detection of
Penicillin-Binding Proteins
-lactam antibiotics with the aid of fluorescence
polarization technology and to monitor a PBP2x protein during purification.
*
Corresponding author. Mailing address: Lilly Research
Laboratories, Infectious Diseases Research; Drop Code 0438, Lilly
Corporate Center, Eli Lilly and Company, Indianapolis, IN 46285-0438. Phone: (317) 276-2040. Fax: (317) 276-1743. E-mail:
Zhao_Genshi{at}Lilly.com.
This article has been cited by other articles:
| Clin. Vaccine Immunol. | Clin. Microbiol. Rev. |
|---|---|
| J. Clin. Microbiol. | ALL ASM JOURNALS |