Fractionation and Characterization of 2',5'-O1igoadenylates by Polyaecrylamide Gel Electrophoresis: An Alternative Method for Assaying 2',5'-O1igoadenylate Synthetase

Mary Beth Miele, Dai Kee Liu, Nancy C. Kan

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

2',5'-Linked oligoadenylates of varying chain lengths (2,5As) are formed from ATP by an interferon (IFN)-induced enzyme, 2',5'-oligoadenylate synthetase (2-5A synthetase). To identify these multiple forms, a method was devised utilizing electrophoretic separation of 32P-labeled 2,5As in a thin 20% polyacrylamide gel containing 7 M urea. A mixture of 2,5As synthesized from rat liver nuclear suspension was fractionated by this method. Each species was eluted from the gel for characterization by specific nucleotidylic enzymes. All major species in the gel were identified, including dimeric, trimeric, and tetrameric forms with either 5' tri- or diphosphates, as well as dephosphorylated species. Thus, in a single step, this method produced a more complete assessment of newly synthesized 2,5As and their degradative products than conventional, multistep DEAE-cellulose chromatography. It also allowed more rapid screening of multiple samples than high-pressure liquid chromatography (HPLC). This method, used to assay 2-5A synthetase induction by IFN-α in human T-cell H9 and CEM-CM3 lines, should be applicable for routine analysis of clinical specimens.

Original languageEnglish (US)
Pages (from-to)33-40
Number of pages8
JournalJournal of Interferon Research
Volume11
Issue number1
DOIs
StatePublished - Feb 1991

All Science Journal Classification (ASJC) codes

  • Immunology
  • Virology

Fingerprint Dive into the research topics of 'Fractionation and Characterization of 2',5'-O1igoadenylates by Polyaecrylamide Gel Electrophoresis: An Alternative Method for Assaying 2',5'-O1igoadenylate Synthetase'. Together they form a unique fingerprint.

  • Cite this