Biochemical and immunological characterization of deoxythymidine kinase of thymidine kinaseless HeLa cells biochemically transformed by herpes simplex virus type.
Thymidine kinase (TK) from herpes simplex virus type 1 (HSV-1) biochemically transformed HeLa cells, purified by affinity chromatography, has been characterized with respect to its electrophoretic mobility, molecular weight, activation energy, substrate specificity, and immunological specificity. TK purified from HSV-1-transformed HeLa cells has the same electrophoretic mobility as TK purified from HeLa cells lytically infected with HSV-1. The sedimentation velocity of purified TK from transformed cells was similar to that previously reported for the lytic enzyme, and its molecular weight was estimated to be 70,000. The activation energy of purified transformed-cell TK was 18.3 kcal/mol. Antiserum prepared against purified HSV-1 TK, although it showed some cross-reactivity, preferentially neutralized homologous TK. The transformed-cell TK antiserum also neutralized the deoxycytidine kinase activity of HSV-1-infected cell extracts but had no effect on deoxycytidine kinase activity of HSV-2-infected cell extract. These results further support the notion that TK acquired by HeLa cells transformed by HSV-1 is of viral and not of cellular origin.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000906:Antibodies; D002846:Chromatography, Affinity; D005796:Genes; D006160:Guanosine Triphosphate / Q000378:metabolism; D006367:HeLa Cells / Q000032:analysis / Q000276:immunology; D009500:Neutralization Tests; D009705:Nucleosides / Q000378:metabolism; D011817:Rabbits; D018139:Simplexvirus / Q000201:enzymology; D013937:Thymidine Kinase / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D014500:Uracil Nucleotides / Q000378:metabolism
D000906:Antibodies; D009705:Nucleosides; D014500:Uracil Nucleotides; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D013937:Thymidine Kinase