title: Synthesis and properties of a nonhydrolyzable adenosine phosphosulfate analog.
abstract: Initial activation of inorganic sulfate for subsequent synthesis of sulfated biomolecules requires the action of ATP-sulfurylase to generate adenosine 5'-phosphosulfate (APS). This activated sulfate intermediate is both chemically labile and susceptible to enzymatic degradation. Consequently, it has not proven useful as a ligand for either purification or characterization of the various APS-utilizing enzymes. For these purposes, a stable analog of APS was required. This paper describes the simple and efficient synthesis and structural confirmation of a nonhydrolyzable APS analog, beta-methylene APS, with an overall molar yield of 40-50%. The method involves nucleophilic substitution of the chlorine moiety of a 5'-chloromethylphosphonate ester of 2',3'-O-isopropylidene adenosine by a sulfite ion. We also report the initial utilization of this compound as an inhibitor in kinetic trials of both ATP-sulfurylase and APS kinase and as an affinity ligand for the purification of these two APS-utilizing enzymes from cartilaginous tissue.
authors.affiliation: Department of Pediatrics, University of Chicago, Illinois 60637.
authors.forename: L
authors.initials: L
authors.lastname: Callahan
authors.forename: K
authors.initials: K
authors.lastname: Ng
authors.forename: D H
authors.initials: DH
authors.lastname: Geller
authors.forename: K
authors.initials: K
authors.lastname: Agarwal
authors.forename: N B
authors.initials: NB
authors.lastname: Schwartz
mesh_terms: D000249:Adenosine Monophosphate / Q000031:analogs & derivatives*; D000250:Adenosine Phosphosulfate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology; D000818:Animals; D002645:Chickens; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D010770:Phosphotransferases / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D017853:Phosphotransferases (Alcohol Group Acceptor); D051381:Rats; D013430:Sulfate Adenylyltransferase / Q000037:antagonists & inhibitors / Q000302:isolation & purification
chemical_list: C060184:beta-methylene adenosine phosphosulfate; D000249:Adenosine Monophosphate; D000250:Adenosine Phosphosulfate; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C019617:adenylylsulfate kinase; D013430:Sulfate Adenylyltransferase