Interactions between T4 phage-coded deoxycytidylate hydroxymethylase and thymidylate synthase as revealed with an anti-idiotypic antibody.
Anti-idiotypic antibodies were used to mimic the binding surface of the T4 bacteriophage deoxycytidylate hydroxymethylase enzyme, providing an immunological probe for protein-protein interactions involving this enzyme. Polyclonal dCMP hydroxymethylase antibodies were affinity-purified and used to generate anti-idiotypic antibodies. The anti-idiotypic serum immunoprecipitated two native viral proteins, deoxycytidylate hydroxymethylase (EC 2.1.2.8) and thymidylate synthase (EC 2.1.1.45), from a sonicated detergent-treated extract of T4-infected Escherichia coli. The anti-anti-dCMP hydroxymethylase antibody was found to be specific in binding to the T4 dTMP synthase, with no detectable affinity for the host dTMP synthase. Previous work in our laboratory has demonstrated the viral dCMP hydroxymethylase and dTMP synthase to be associated in a deoxyribonucleotide synthetase enzyme complex. Our current approach, using anti-idiotypic antibodies as probes for protein-protein interactions, and complementary studies involving dCMP hydroxymethylase enzyme affinity columns indicate a direct association between bacteriophage T4 dCMP hydroxymethylase and dTMP synthase.
Young
J P
JP
Department of Biochemistry and Biophysics, Oregon State University, Corvallis 97331-6503.
Mathews
C K
CK
D000888:Antibodies, Anti-Idiotypic; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral; D019877:Hydroxymethyl and Formyl Transferases; D011233:Precipitin Tests; D013604:T-Phages / Q000201:enzymology*; D013940:Thymidylate Synthase / Q000235:genetics / Q000378:metabolism*; D014166:Transferases / Q000235:genetics / Q000378:metabolism*
D000888:Antibodies, Anti-Idiotypic; D014166:Transferases; D013940:Thymidylate Synthase; D019877:Hydroxymethyl and Formyl Transferases; C100226:deoxycytidylate hydroxymethyltransferase