Identification of tyrosylprotein sulfotransferase in rat gastric mucosa.
An enzyme activity which catalyzes the transfer of the sulfate group from 3'-phosphoadenosine 5'-phosphosulfate (PAPS) to poly-Glu6,Ala3,Tyr1 (EAY; M(r) 47,000) has been demonstrated in the antral and body mucosa of the rat stomach. The distribution of this tyrosylprotein sulfotransferase was similar to that of the Golgi marker enzyme, glycoprotein sulfotransferase, and its activity from body mucosa was 23% higher than that from the antrum. The optimum for tyrosylprotein sulfotransferase activity was obtained at pH 6.8, in the presence of 0.5% Triton X-100, 20 mmol/l MnCl2, 50 mmol/l NaF, 2 mmol/l 5'-AMP, and 1 mmol/l DTT, whereas Ca2+, Mg2+, Cu2+, Zn2+, EDTA, NEM, NaCl and Na2SO4 were inhibitory. The apparent Km of the sulfotransferase for EAY was 1.5 x 10(-6) mol/l and for PAPS 0.75 x 10(-6) mol/l. The enzyme was 28 times less susceptible to 2,6-dichloro-4-nitrophenol inhibition as compared to that required for phenol sulfotransferase inhibition. The tyrosine sulfation by the tyrosylprotein sulfotransferase was independent of the sulfation of carbohydrate residues in mucous glycoproteins and glycolipids, thus indicating that the identified sulfotransferase is specific for sulfation of the tyrosyl residues in the peptide core.
Kasinathan
C
C
Research Center, University of Medicine and Dentistry of New Jersey, Newark.
Sundaram
P
P
Slomiany
B L
BL
Slomiany
A
A
D000818:Animals; D002413:Cations, Divalent; D002712:Chlorides; D005753:Gastric Mucosa / Q000201:enzymology*; D007700:Kinetics; D008297:Male; D008345:Manganese / Q000494:pharmacology; D017895:Manganese Compounds; D051381:Rats; D017207:Rats, Sprague-Dawley; D012969:Sodium Fluoride / Q000494:pharmacology; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology; D015238:Sulfotransferases / Q000302:isolation & purification / Q000378:metabolism*
D002413:Cations, Divalent; D002712:Chlorides; D017895:Manganese Compounds; D008345:Manganese; D012969:Sodium Fluoride; D015238:Sulfotransferases; C042860:glycoprotein sulfotransferase; C039079:protein-tyrosine sulfotransferase; D000254:Sodium-Potassium-Exchanging ATPase; C025340:manganese chloride