Switch to
Predicate | Object |
---|---|
rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
|
pubmed:dateCreated |
1989-8-10
|
pubmed:abstractText |
A solution of propionic acid, 1 M ammonium hydroxide, and isopropyl alcohol (45/17.5/17.5, v/v) was the ascending solvent in the separation of phosphotyrosine, phosphothreonine, and phosphoserine by thin-layer chromatography. The immobile phase was cellulose. The relative migrations were 0.44, 0.38, and 0.2, respectively. A previously described thin-layer system consisting of isobutyric acid and 0.5 M ammonium hydroxide (50/30, v/v) gave very similar relative migrations. To determine the usefulness of thin-layer chromatography in phosphoamino acid analysis, the propionic acid/ammonium hydroxide/isopropyl alcohol solution was used to characterize phosphorylated residues in a plasma membrane protein which is a substrate for the insulin receptor kinase, in insulin receptor phosphorylated histone H2B, and in an in vivo phosphorylated 90000-Da protein from IM9 cells. 32P-labeled proteins were separated by dodecyl sulfate-gel electrophoresis, digested with trypsin, and then hydrolyzed with 6 N HCl, 2 h, 110 degrees C. Following thin-layer chromatography of the hydrolyzates and autoradiography, phosphotyrosine was detected in insulin receptor substrates, and phosphoserine and phosphothreonine were found in the in vivo-phosphorylated protein. This study supports previous reports about the practicality of thin-layer chromatography in phosphoamino acid analysis and it demonstrates that a propionic acid, ammonium hydroxide, isoprophyl alcohol solution may be a useful ascending solvent mixture for this purpose.
|
pubmed:language |
eng
|
pubmed:journal | |
pubmed:citationSubset |
IM
|
pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Phosphoproteins,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphorus Radioisotopes,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphoserine,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphothreonine,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphotyrosine,
http://linkedlifedata.com/resource/pubmed/chemical/Serine,
http://linkedlifedata.com/resource/pubmed/chemical/Threonine,
http://linkedlifedata.com/resource/pubmed/chemical/Tyrosine
|
pubmed:status |
MEDLINE
|
pubmed:month |
Feb
|
pubmed:issn |
0003-2697
|
pubmed:author | |
pubmed:issnType |
Print
|
pubmed:day |
15
|
pubmed:volume |
177
|
pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
|
pubmed:pagination |
138-43
|
pubmed:dateRevised |
2006-11-15
|
pubmed:meshHeading |
pubmed-meshheading:2472754-Animals,
pubmed-meshheading:2472754-Autoradiography,
pubmed-meshheading:2472754-Chromatography, Thin Layer,
pubmed-meshheading:2472754-Humans,
pubmed-meshheading:2472754-Hydrolysis,
pubmed-meshheading:2472754-Phosphoproteins,
pubmed-meshheading:2472754-Phosphorus Radioisotopes,
pubmed-meshheading:2472754-Phosphorylation,
pubmed-meshheading:2472754-Phosphoserine,
pubmed-meshheading:2472754-Phosphothreonine,
pubmed-meshheading:2472754-Phosphotyrosine,
pubmed-meshheading:2472754-Rats,
pubmed-meshheading:2472754-Serine,
pubmed-meshheading:2472754-Threonine,
pubmed-meshheading:2472754-Tyrosine
|
pubmed:year |
1989
|
pubmed:articleTitle |
Thin-layer chromatography can resolve phosphotyrosine, phosphoserine, and phosphothreonine in a protein hydrolyzate.
|
pubmed:affiliation |
Department of Medical Biochemistry, Faculty of Medicine, University of Calgary, Alberta, Canada.
|
pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|