Rat Chondrosarcoma ATP Sulfurylase and Adenosine 5'-Phosphosulfate Kinase Reside on a Single Bifunctional Protein

The sulfate-activation pathway consists of the sequential action of ATP sulfurylase (ATP: sulfate adenylyltransferase, EC 2.7.7.4) and adenosine 5'-phosphosulfate kinase ATP:adenylylsulfate 3'-phosphotransferase, EC 2.7.1.25). Both sulfurylase and kinase from rat chondrosarcoma were copuri...

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Bibliographic Details
Published in:Biochemistry (Easton) 1994-05, Vol.33 (19), p.5920-5925
Main Authors: Lyle, Stephen, Stanczak, Jeffrey, Ng, Kian, Schwartz, Nancy B
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Antibodies
Chondrosarcoma - enzymology
Chromatography, Affinity
Chromatography, Gel
Electrophoresis, Polyacrylamide Gel
Molecular Sequence Data
Phosphotransferases (Alcohol Group Acceptor) - chemistry
Phosphotransferases (Alcohol Group Acceptor) - immunology
Phosphotransferases (Alcohol Group Acceptor) - isolation & purification
Phosphotransferases (Alcohol Group Acceptor) - metabolism
Rats
Sulfate Adenylyltransferase - chemistry
Sulfate Adenylyltransferase - immunology
Sulfate Adenylyltransferase - isolation & purification
Sulfate Adenylyltransferase - metabolism
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Summary:The sulfate-activation pathway consists of the sequential action of ATP sulfurylase (ATP: sulfate adenylyltransferase, EC 2.7.7.4) and adenosine 5'-phosphosulfate kinase ATP:adenylylsulfate 3'-phosphotransferase, EC 2.7.1.25). Both sulfurylase and kinase from rat chondrosarcoma were copurified through substrate affinity chromatography using stable analogs of APS (adenosine 5'-phosphosulfate) and PAPS (3'-phosphoadenosine 5'-phosphosulfate). A 56-kDa protein, containing both activities, was then purified to apparent homogeneity through reversed-phase chromatography and observed on denaturing polyacrylamide gels. The molecular mass of the native active unit containing both activities in the purified preparation corresponded to approximately 60 kDa by analytical gel filtration. Coincident binding and elution of ATP sulfurylase and APS kinase by immunoaffinity chromatography, using polyclonal serum generated against the 56-kDa protein, also demonstrated that the enzyme contains both activities. Lastly, a single N-terminal amino acid sequence was obtained from the 56-kDa band isolated by gel electrophoresis. These results all suggest that ATP sulfurylase and APS kinase from rat chondrosarcoma residue on a single bifunctional protein.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi00185a032