3'-Phosphoadenosine 5'-phosphosulfate biosynthesis and the sulfation of cholecystokinin by the tyrosylprotein-sulfotransferase in rat brain tissue.
This article resumes the work we have accomplished in the past few years. Cholecystokinin sulfation is an important post-translational modification necessary for the biological activity of this peptide hormone. The tyrosyl protein sulfotransferase (TPST) activity from rat cerebral cortex was characterized. TPST activity is most probably responsible for the endogenous sulfation of CCK. TPST reaction kinetic properties were studied using radiolabeled 3'-phosphoadenosine 5'-phosphosulfate (PAPS) and the non-sulfated peptide acceptor terbutyloxycarbonyl-cholecystokinin octapeptide (BocCCK-8(ns)) as substrates, and brain microsomes as the enzyme source. The BocCCK-8 sulfating reaction data is consistent with the idea that TPST forward reaction follows an ordered Bi Bi mechanism. PAPS biosynthesis and availability was studied in slices from rat cerebral cortex incubated in the presence of [35S]sulfate. There is a rapid and dynamic turnover of the steady-state level of PAPS in brain cells which is decreased by depolarizing agents such as potassium, veratridine and glutamate. Furthermore, the presence of a membrane-bound PAPS biosynthesis inhibitor was observed. These results are discussed in view of the biological importance that the cell sulfating pathways might play in nerve cell activity.
Laboratoire de Neurobiologie et Pharmacologie, Unite 109, Centre Paul Broca de l'Inserm, Paris, France.
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Vargas
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Frerot
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Brion
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Trung Tuong
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Lafitte
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Gulat-Marnay
D000818:Animals; D002540:Cerebral Cortex / Q000201:enzymology*; D066298:In Vitro Techniques; D010724:Phosphoadenosine Phosphosulfate / Q000096:biosynthesis* / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D051381:Rats; D012844:Sincalide / Q000378:metabolism*; D013431:Sulfates / Q000378:metabolism; D015238:Sulfotransferases / Q000378:metabolism*
D013431:Sulfates; D010724:Phosphoadenosine Phosphosulfate; D015238:Sulfotransferases; C039079:protein-tyrosine sulfotransferase; D012844:Sincalide