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In Situ Autoradiographic Detection of Folylpolyglutamate Synthetase Activity by Daniel J. Sussman; Gregory Milman; Charles Osborne; Barry Shane is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is In Situ Autoradiographic Detection of Folylpolyglutamate Synthetase Activity about?

The enzyme folylpolyglutamate synthetase (FPGS) catalyzes the conversion of folate (pteroylmonoglutamate) to the polyglutamate forms (pteroylpolyglutamates) that are required for folate retention by mammalian cells. A rapid in situ autoradiographic assay for FPGS was developed which is based on the folate cofactor requirement of thymidylate synthase. Chinese hamster AUX 91 mutant cells lack FPGS activity and are unable to accumulate folate. As a result, the conversion of [6-3H]deoxyuridine to thymidine via the thymidylate synthase reaction is impaired in AUX B I cells and no detectable label is incorporated into DNA. In contrast, FPGS in wild-type Chinese hamster CHO cells causes folate retention and enables the incorporation of [6-3H]deoxyuridine into DNA. Incorporation may be detected by autoradiography of monolayer cultures or of colonies replica plated onto polyester discs. Introduction of Escherichia coli FPGS into AUX BI cells restores the activity of the thymidylate synthase pathway and demonstrates that the E. coli FPGS enzyme can provide pteroylpolygmtamates which function in mammalian cells. cc' 1986 Academic Press. Inc.

Who reads In Situ Autoradiographic Detection of Folylpolyglutamate Synthetase Activity?

Author

Daniel J. Sussman; Gregory Milman; Charles Osborne; Barry Shane

Publisher

Elsevier Science; Elsevier ; Elsevier Inc.; Elsevier BV (ISSN 0003-2697)

Published

1986

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)