Paper disk assay for glycosaminoglycan sulfotransferases

Sugahara, K; Ishii, T; Yamashina, I

doi:10.1016/0003-2697(87)90591-4

Title: Paper disk assay for glycosaminoglycan sulfotransferases

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Abstract

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.

Authors:: Sugahara, K; Ishii, T; Yamashina, I

Publication Date:: Sun Nov 01 00:00:00 EST 1987

Research Org.:: Kyoto Univ. (Japan)

OSTI Identifier:: 5272113

Resource Type:: Journal Article

Journal Name:: Anal. Biochem.; (United States)

Additional Journal Information:: Journal Volume: 166:2

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; 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

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                    Sugahara, K, Ishii, T, and Yamashina, I. Paper disk assay for glycosaminoglycan sulfotransferases.  United States: N. p., 1987. 
        Web.  doi:10.1016/0003-2697(87)90591-4.

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                    Sugahara, K, Ishii, T, & Yamashina, I. Paper disk assay for glycosaminoglycan sulfotransferases.  United States.  https://doi.org/10.1016/0003-2697(87)90591-4

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                    Sugahara, K, Ishii, T, and Yamashina, I. 1987.  
        "Paper disk assay for glycosaminoglycan sulfotransferases".  United States.  https://doi.org/10.1016/0003-2697(87)90591-4.

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@article{osti_5272113,

  title        = {Paper disk assay for glycosaminoglycan sulfotransferases},

  author       = {Sugahara, K and Ishii, T and Yamashina, I},

  abstractNote = {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.},

  doi          = {10.1016/0003-2697(87)90591-4},

  url          = {https://www.osti.gov/biblio/5272113},
  journal      = {Anal. Biochem.; (United States)},
number       = ,

  volume       = 166:2,

  place        = {United States},

  year         = {Sun Nov 01 00:00:00 EST 1987},

  month        = {Sun Nov 01 00:00:00 EST 1987}

}

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