MolPharm xPharm- The Comprehensive Pharmacology Reference

Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text (PDF)
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
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
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Morris, G.
Right arrow Articles by Lazo, J. S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Morris, G.
Right arrow Articles by Lazo, J. S.

Neutralization of bleomycin hydrolase by an epitope-specific antibody

G Morris, JS Mistry, JP Jani, JE Mignano, SM Sebti and JS Lazo

Department of Pharmacology, University of Pittsburgh, School of Medicine, Pennsylvania 15261.

Bleomycin hydrolase (BH) is a cysteine proteinase that terminates the pharmacological action of bleomycin (BLM). Amino acid sequence data obtained from a tryptic digest fragment of purified rabbit lung BH were used to synthesize a 14-amino acid peptide (LAVLEQEPIVLPAK; BHP14), which was conjugated to horseshoe crab hemocyanin and used to produce rabbit antiserum that was immunoreactive to both BHP14 and rabbit BH. Anti-BHP14 binding to BHP14 could be competitively blocked by the presence of either BHP14 or BH. Anti-BHP14 recognized both purified rabbit liver BH and postmicrosomal fraction from rabbit liver on Western blot, as a single band of M(r) approximately 48,000. Anti-BHP14 inhibited, in a concentration-dependent manner, BH activity in rabbit liver cytosolic fractions, as measured by deamido-BLM A2 formation. Thus, we have generated an epitope-specific neutralizing antibody to rabbit BH, which can block the metabolism of BLM by homogenates from rabbit tissue. These results suggest that the LAVLEQEPIVLPA epitope of rabbit BH is involved in the metabolism of BLM or is topologically near the active site. Furthermore, a BLM-resistant squamous carcinoma (C- 10E) exhibited slightly more immunoreactivity, by enzyme-linked immunosorbent assay, to anti-BHP14 than did the parental A-253 cells, and a partially revertant (C-10E ND) cell line had intermediate anti- BHP14 binding. BH activity in these cell lines was in the same rank order as antibody binding, but differences in immunoreactivity were less than differences in enzymatic activity. Our epitope-specific neutralizing antibody should be useful in the further characterization of BH.

Volume 42, Issue 1, pp. 57-62, 07/01/1992
Copyright © 1992 by American Society for Pharmacology and Experimental Therapeutics




This article has been cited by other articles:


Home page
 JCOHome page
E. C. de Haas, N. Zwart, C. Meijer, J. Nuver, H. M. Boezen, A. J.H. Suurmeijer, H. J. Hoekstra, G. van der Steege, D. Th. Sleijfer, and J. A. Gietema
Variation in Bleomycin Hydrolase Gene Is Associated With Reduced Survival After Chemotherapy for Testicular Germ Cell Cancer
J. Clin. Oncol., April 10, 2008; 26(11): 1817 - 1823.
[Abstract] [Full Text] [PDF]


Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
All ASPET Journals Molecular Pharmacology Pharmacological Reviews
Molecular Interventions Drug Metabolism and Disposition

Copyright © 1992 by the American Society for Pharmacology and Experimental Therapeutics