An Enzymatic Procedure for the Preparation and Purification of 3′-Phosphoadenosine 5′-Phospho- [35S]Sulfate ([35S]PAPS): Applications in Syntheses of 8-Azido and 8-Bromo Derivatives of [35S]PAPS
✍ Scribed by K. Shailubhai; R.K. Singh; J.J. Schmuke; G.S. Jacob
- Publisher
- Elsevier Science
- Year
- 1996
- Tongue
- English
- Weight
- 165 KB
- Volume
- 243
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
This paper describes a rapid and an efficient procethe ''active sulfate'' used as a primary donor of sulfuryl dure for the enzymatic synthesis of 3 -phosphoadenosgroups (0SO 3 ) in biological systems (1). Sulfotransferine 5-phospho[ 35 S]sulfate ([ 35 S]PAPS). [ 35 S]PAPS was ases are a family of enzymes that catalyze the transfer synthesized by incubating ATP and a carrier-free [ 35 S]of the sulfuryl group from PAPS to a variety of acceptor Na 2 35 SO 4 with ATP sulfurylase, a recombinant APS kisubstrates including xenobiotics, proteins, glycolipids, nase and inorganic pyrophosphatase. The transfer of steroids, and glycosaminoglycans (2-6). Since the dis-35 SO 4 group from [ 35 S]Na 2 SO 4 to [ 35 S]PAPS proceeded covery of active sulfate, numerous chemical and enzymore efficiently in the presence of an ATP-regeneratmatic attempts have been made to synthesize both ing system composed of pyruvate kinase and phosadenosine phosphosulfate (APS) and PAPS. In biologiphoenol pyruvate. About 90% of the radioactivity prescal systems, PAPS is synthesized through the coordient in the starting material [ 35 S]Na 2 SO 4 was transnated action of three enzymes as outlined below (7).
ferred to [ 35 S]PAPS within a 2-h reaction incubation. The reaction products were applied to a Mono Q column, and [ 35 S]PAPS was eluted by a step-wise gradient Academic Press, Inc.