Partial purification and characterization of 3'-phosphoadenylylsulfate:keratan sulfate sulfotransferases

Two 3'-phosphoadenylylsulfate:keratan sulfate sulfotransferases were purified 600-fold and 340-fold, respectively, from isolated bovine cornea cells. Sulfotransferase I exhibited an apparent Mr = 220,000, whereas an Mr = 140,000 was calculated for sulfotransferase II. The final preparations wer...

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Bibliographic Details
Published in:The Journal of biological chemistry 1984-10, Vol.259 (19), p.11771-11776
Main Authors: RUTER, E.-R, KRESSE, H
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Animals
Biological and medical sciences
Carbohydrate Sulfotransferases
Chromatography, High Pressure Liquid
Cornea - enzymology
Enzymes and enzyme inhibitors
Fundamental and applied biological sciences. Psychology
Hydrogen-Ion Concentration
Keratan Sulfate - metabolism
Magnetic Resonance Spectroscopy
Molecular Weight
Phosphoadenosine Phosphosulfate - metabolism
Rats
Sulfates - metabolism
Sulfotransferases
Sulfurtransferases - isolation & purification
Transferases
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Summary:Two 3'-phosphoadenylylsulfate:keratan sulfate sulfotransferases were purified 600-fold and 340-fold, respectively, from isolated bovine cornea cells. Sulfotransferase I exhibited an apparent Mr = 220,000, whereas an Mr = 140,000 was calculated for sulfotransferase II. The final preparations were both devoid of chondroitin sulfate sulfotransferase activity. The position of sulfation was determined by proton nuclear magnetic resonance spectroscopy. Sixty per cent of the sulfate ester groups formed by sulfotransferase I were linked to the C-6 atom of galactosyl residues, the other ones to the C-6 atom of N-acetylglucosamine. Sulfotransferase II showed a different specificity: 23% of the newly formed sulfate ester groups were on galactosyl and 77% on N-acetylglucosaminyl residues. Both sulfotransferase preparations acted in a cooperative manner. In the presence of both sulfotransferases, the incorporation of [35S]sulfate into keratan sulfate was up to 75% higher than could be expected from the sum of individual activities. From the specific radioactivities of the oligosaccharides produced by digestion with endo-beta-galactosidase, it was also concluded that both enzyme species reacted best with keratan sulfate segments exhibiting a relatively high degree of sulfation.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(20)71278-0