Synthesis of a new 8-spin-labeled analog of adenosine 5'-phosphate and its interaction with AMP nucleosidase.

W. E. DeWolf, Vern L. Schramm

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The synthesis of a new 8-spin-labeled analog of AMP, 8-[[[(2,2,5,5-tetramethyl-1-oxy-3-pyrrolidinyl)carbamoyl]methyl]thio]adenosine 5'-phosphate (8-slAMP), is described. The procedure is facile and results in high yields. 8-slAMP is a competitive inhibitor of AMP nucleosidase with a Ki of 19 microM as compared to a Km of 100 microM for AMP. The analog is not a substrate for the enzyme and does not displace MgATP2- from the allosteric sites under the usual assay conditions. The EPR spectrum of the bound spin probe reveals a highly immobilized nitroxide group. Binding studies with 8-slAMP at 8 degrees C indicate three independent binding sites (Kd = 1.4 microM) per molecule of enzyme (Mr = 320,000). These properties make 8-slAMP a good spin probe for AMP nucleosidase. The analog may also be useful for other proteins known or suspected of binding AMP analogs in a syn conformation.

Original languageEnglish (US)
Pages (from-to)6215-6217
Number of pages3
JournalJournal of Biological Chemistry
Volume254
Issue number14
StatePublished - Jul 25 1979
Externally publishedYes

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AMP nucleosidase
Adenosine Monophosphate
Allosteric Site
Enzymes
Paramagnetic resonance
Conformations
Assays
Binding Sites
Molecules
Substrates
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Synthesis of a new 8-spin-labeled analog of adenosine 5'-phosphate and its interaction with AMP nucleosidase. / DeWolf, W. E.; Schramm, Vern L.

In: Journal of Biological Chemistry, Vol. 254, No. 14, 25.07.1979, p. 6215-6217.

Research output: Contribution to journalArticle

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