Paper disk assay for glycosaminoglycan sulfotransferases
A method is described for the assay of sulfotransferases, which transfer sulfate from 3'-phosphoadenosine-5'-phosphosulfate (PAPS) to glycosaminoglycan acceptors. Following the sulfation reactions, the (/sup 35/S)sulfate-labeled products are precipitated and then separated from a sulfate donor ((/sup 35/S)PAPS) and its degradation products by a paper disk method, and then the radioactivity remaining on the paper disk is subsequently determined by liquid scintillation counting. The rapidity and simplicity of the method are advantageous for multiple assays and have allowed us to establish assay conditions for serum sulfotransferases which introduce sulfate at position 6 of the internal N-acetylgalactosamine units of chondroitin, position 2 (amino group) of the glucosamine units of heparan sulfate and sugar units of keratan sulfate, respectively. The assay method will be applicable with modification to the assay of other glycosaminoglycan sulfotransferases and glycoprotein sulfotransferases.
- Research Organization:
- Kyoto Univ. (Japan)
- OSTI ID:
- 5272113
- Journal Information:
- Anal. Biochem.; (United States), Vol. 166:2
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
TRANSFERASES
BIOASSAY
BUFFERS
CHICKENS
GLUCOPROTEINS
SCINTILLATION COUNTING
SULFATES
SULFUR 35
TRACER TECHNIQUES
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIRDS
CARBOHYDRATES
COUNTING TECHNIQUES
DAYS LIVING RADIOISOTOPES
ENZYMES
EVEN-ODD NUCLEI
FOWL
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
NUCLEI
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PROTEINS
RADIOISOTOPES
SACCHARIDES
SULFUR COMPOUNDS
SULFUR ISOTOPES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques