Linked Life Data

8621720

Source:http://linkedlifedata.com/resource/pubmed/id/8621720

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6
pubmed:dateCreated
1996-6-19
pubmed:abstractText
Photoaffinity labeling with 2-azidoadenosine 3', 5'-[5'-32P]bisphosphate was used to identify and characterize adenosine 3',5'-bisphosphate-binding proteins in human liver cytosol and recombinant sulfotransferase proteins. The sulfotransferases investigated in these studies were the human phenol sulfotransferases, HAST1, -3, and -4, dehydroepiandrosterone sulfotransferase, and estrogen sulfotransferase. The cDNAs for these enzymes have been previously cloned and expressed in COS-7 cells or Escherichia coli. Photoaffinity labeling of all proteins was highly dependent on UV irradiation, was protected by co-incubation with unlabeled adenosine 3',5'-bisphosphate and phosphoadenosine phosphosulfate, and reached saturation at concentrations above 10 microM. To verify that the 31 35-kDa photolabeled proteins were indeed sulfotransferases, specific antibodies known to recognize human sulfotransferases were used for Western blot analyses of photolabeled proteins. It was shown unequivocally that the proteins in the 31-35-kDa region recognized by the antibodies also photoincorporated 2-azidoadenosine 3',5'-[5'-32P]bisphosphate. This is the first application of photoaffinity labeling with 2-azidoadenosine 3',5'-[5'-32P]bisphosphate for the characterization of recombinant human sulfotransferases. Photoaffinity labeling will be also useful in the purification and functional identification of other adenosine 3',5'-bisphosphate-binding proteins and to determine amino acid sequences at or near their active sites.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
9
pubmed:volume
271
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
3195-9
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:8621720-Adenosine Diphosphate, pubmed-meshheading:8621720-Affinity Labels, pubmed-meshheading:8621720-Animals, pubmed-meshheading:8621720-Arylsulfotransferase, pubmed-meshheading:8621720-Autoradiography, pubmed-meshheading:8621720-Azides, pubmed-meshheading:8621720-Blotting, Western, pubmed-meshheading:8621720-Cell Line, pubmed-meshheading:8621720-Cercopithecus aethiops, pubmed-meshheading:8621720-Cloning, Molecular, pubmed-meshheading:8621720-Cytosol, pubmed-meshheading:8621720-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:8621720-Endoplasmic Reticulum, pubmed-meshheading:8621720-Escherichia coli, pubmed-meshheading:8621720-Humans, pubmed-meshheading:8621720-Liver, pubmed-meshheading:8621720-Phosphorus Radioisotopes, pubmed-meshheading:8621720-Rats, pubmed-meshheading:8621720-Recombinant Proteins, pubmed-meshheading:8621720-Sulfotransferases, pubmed-meshheading:8621720-Transfection
pubmed:year
1996
pubmed:articleTitle
Photoaffinity labeling of human recombinant sulfotransferases with 2-azidoadenosine 3',5'-[5'-32P]bisphosphate.
pubmed:affiliation
Department of Internal Medicine, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't