Subcellular and developmental studies of the tyrosyl protein sulfotransferase in rat brain.
1. Tyrosyl protein sulfotransferase (TPS) activity in the newborn and mature rat brain was studied using the cholecystokinin derivative terbutyloxycarbonyl-Asp-Tyr-Met-Gly-Trp-Met-Asp-PheNH2, BocCCK-8(ns), as the peptide substrate. 2. TPS activity was enriched 4 times in the microsomal and synaptic vesicular enriched fractions of rat cerebral cortex. 3. CCK-8 content, in the subcellular fractions and the peptide sulfation activity distribution was in accord with the hypothesis that tyrosyl protein sulfotransferase plays a key role in the maturation process of bioactive CCK. 4. TPS activity measured in membranes from newborn brain was 2.5 times higher than the activity observed in the mature brain membranes with a Vmax = 0.83 +/- 0.05 and 0.31 +/- 0.02 respectively. The apparent KM for the sulfate donor, 3'-phosphoadenosine 5'-phosphosulfate (PAPS), was similar, 94 +/- 4 nM and 90 +/- 6 nM and the KM for the peptide substrate, BocCCK-8(ns), was 234 +/- 16 microM and 160 +/- 12 microM in the newborn and adult brain membranes respectively. 5. TPS activity reached normal mature values within 20 days of age. 6. These data support the idea that tyrosyl protein sulfation is an important process in the secretion mechanism and in the CCK maturation.
Dam Trung Tuong
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M
Unité de Neurobiologie et Pharmacologie (U. 109) de l'INSERM, Centre Paul Broca, Paris, France.
Vargas
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F
Brion
F
F
Frerot
O
O
D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000201:enzymology*; D002540:Cerebral Cortex / Q000201:enzymology; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D008861:Microsomes / Q000201:enzymology; D008969:Molecular Sequence Data; D051381:Rats; D017207:Rats, Sprague-Dawley; D012844:Sincalide / Q000031:analogs & derivatives / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology; D015238:Sulfotransferases / Q000378:metabolism*; D013572:Synaptic Vesicles / Q000201:enzymology
C061447:tert-butyloxycarbonylcholecystokinin-8; D015238:Sulfotransferases; C039079:protein-tyrosine sulfotransferase; D012844:Sincalide