Synthesis and properties of a nonhydrolyzable adenosine phosphosulfate analog.

L Callahan, K Ng, D H Geller, K Agarwal, N B Schwartz

Department of Pediatrics, University of Chicago, Illinois 60637.

Analytical biochemistry 1989 Feb 15

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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.

Citation

L Callahan, K Ng, D H Geller, K Agarwal, N B Schwartz. Synthesis and properties of a nonhydrolyzable adenosine phosphosulfate analog. Analytical biochemistry. 1989 Feb 15;177(1):67-71