Open AccessComparative complement selection in bacteria enables screening for lead compounds targeted to a purine salvage enzyme of parasites
Author(s) -
Ann E. Eakin,
René Nieves-Alicea,
Rafael Tosado-Acevedo,
Michael C. Chin,
C C Wang,
Sydney P. Craig
Publication year - 1995
Publication title -
antimicrobial agents and chemotherapy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.07
H-Index - 259
eISSN - 1070-6283
pISSN - 0066-4804
DOI - 10.1128/aac.39.3.620
Subject(s) - biology , bacteria , escherichia coli , hypoxanthine guanine phosphoribosyltransferase , recombinant dna , complementation , plasmid , nucleotide salvage , schistosoma mansoni , enzyme , biochemistry , microbiology and biotechnology , genetics , dna , gene , nucleotide , schistosomiasis , zoology , mutant , helminths , phenotype
Expression plasmids encoding the hypoxanthine phosphoribosyltransferases (HPRTs) of Plasmodium falciparum, Schistosoma mansoni, Tritrichomonas foetus, and Homo sapiens were subcloned into genetically deficient Escherichia coli that requires complementation by the activity of a recombinant HPRT for growth on semidefined medium. Fifty-nine purine analogs were screened for their abilities to inhibit the growth of these bacteria. Several compounds that selectively altered the growth of the bacteria complemented by the malarial, schistosomal, or tritrichomonal HPRT compared with the growth of bacteria expressing the human enzyme were identified. These results demonstrate that the recombinant approach to screening compounds by complement selection in a comparative manner provides a rapid and efficient method for the identification of new lead compounds selectively targeted to the purine salvage enzymes of parasites.