Development and characterization of a rapid screening assay for identifying antipneumocystis agents

A. Martinez, J. A. Kovacs

Research output: Contribution to journalArticlepeer-review


We developed a rapid assay for screening of compounds with potential antipneumocystis activity on the basis of incorporation of [35S]methionine into proteins newly synthesized by Pneumocystis carinii. Unambiguous evidence that P. carinii synthesizes proteins in vitro was provided by immunoprecipitation studies demonstrating the incorporation of [35S]methionine into the major surface glycoprotein. Treatment with two clinically active antipneumocystis agents, atovaquone (10-4 M) or pentamidine (10-4 M), prevented this incorporation. Total [35S]methionine incorporation paralleled incorporation into the major surface glycoprotein, permitting rapid assessment of anti-P. carinii activity by scintillation counting. Treatment with pentamidine (1 x 10-4 M), atovaquone, trimethoprim (1 x 10-4 M)-sulfamethoxazole (7.9 x 10-4 M), piritrexim (1 x 10-7 M), RO11-8958 (1 x 10-4 M), and amphotericin B (1 μg/ml) resulted in a greater than 67% inhibition (P < 0.05) of [35S]methionine incorporation. No decrease in [35S]methionine incorporation was seen with dapsone (10-5 M), trimethoprim (10-4 M), recombinant mouse tumor necrosis factor (500 ng/ml), or gamma interferon. This rapid in vitro assay should be a useful adjunct in the development of new antipneumocystis agents.

Original languageEnglish (US)
Pages (from-to)1674-1678
Number of pages5
JournalAntimicrobial agents and chemotherapy
Issue number8
StatePublished - 1993

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)
  • Infectious Diseases


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