Human cationic trypsinogen is sulfated on Tyr154.
The crystal structure of human pancreatic cationic trypsin showed the chemical modification of Tyr154, which was originally described as phosphorylation [Gaboriaud C, Serre L, Guy-Crotte O, Forest E & Fontecilla-Camps JC (1996) J Mol Biol259, 995-1010]. Here we report that Tyr154 is sulfated, not phosphorylated. Cationic and anionic trypsinogens were purified from human pancreatic juice and subjected to alkaline hydrolysis. Modified tyrosine amino acids were separated on a Dowex cation-exchange column and analyzed by thin layer chromatography. Both human cationic and anionic trypsinogens contained tyrosine sulfate, but no tyrosine phosphate, whereas bovine trypsinogen contained neither. Furthermore, incorporation of [(35)S]SO(4) into human cationic trypsinogen transiently expressed by human embryonic kidney 239T cells was demonstrated. Mutation of Tyr154 to Phe abolished radioactive sulfate incorporation, confirming that Tyr154 is the site of sulfation in cationic trypsinogen. Sulfated pancreatic cationic trypsinogen exhibited faster autoactivation than a nonsulfated recombinant form, suggesting that tyrosine sulfation of trypsinogens might enhance intestinal digestive zymogen activation in humans. Finally, sequence alignment revealed that the sulfation motif is only conserved in primate trypsinogens, suggesting that typsinogen sulfation is absent in other vertebrates.
Sahin-Tóth
Miklós
M
Department of Molecular and Cell Biology, Boston University, Goldman School of Dental Medicine, Boston, MA, USA. miklos@bu.edu
Kukor
Zoltán
Z
Nemoda
Zsófia
Z
D000818:Animals; D002412:Cations / Q000737:chemistry / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D002855:Chromatography, Thin Layer; D004586:Electrophoresis / Q000379:methods; D004789:Enzyme Activation; D006801:Humans; D006868:Hydrolysis; D010189:Pancreatic Juice / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D013455:Sulfur / Q000378:metabolism; D013457:Sulfur Compounds / Q000493:pharmacokinetics; D013460:Sulfur Isotopes / Q000493:pharmacokinetics; D014362:Trypsinogen / Q000737:chemistry* / Q000378:metabolism*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000302:isolation & purification
D002412:Cations; D013457:Sulfur Compounds; D013460:Sulfur Isotopes; C005662:tyrosine O-sulfate; D014443:Tyrosine; D013455:Sulfur; D014362:Trypsinogen