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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
52
pubmed:dateCreated
1995-1-30
pubmed:abstractText
The transforming gene v-crk found in CT10 and ASV-1 avian sarcoma viruses induces marked phosphorylation of several proteins in cells expressing p47v-crk (v-Crk). In this work, the main tyrosine-phosphorylated proteins in ASV-1-infected chicken cells and v-crk-transfected rat cells were characterized biochemically. Both these proteins have a molecular mass of about 130 kDa and are tightly associated with v-Crk in vivo. Two-dimensional gel electrophoresis revealed that they are both essentially single proteins (p130) with modifications that result in a broad spot in an acidic region. The broad band of semi-purified p130 became sharp at an elevated position in the gel upon treatment with orthovanadate in vivo or with c-Src kinase produced using a baculovirus vector in vitro, whereas it shifted at a lower position upon treatment with alkaline phosphatase in vitro. These results suggest multiple phosphorylation states of p130, which result in a broad band of p130. Two procedures of immunoaffinity purification were used to purify p130 from 3Y1 cells transfected with v-crk. Approximately 30 pmol of purified p130 was obtained in an immobilized form on a filter starting from 3 x 10(10) cells. Peptide mapping of p130 digested in situ by peptidase revealed that the purity and quantity of the final material were enough for peptide sequencing. Several stretches of partial amino acid sequences were determined, and they indicated that p130 is a novel protein.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
30
pubmed:volume
269
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
32740-6
pubmed:dateRevised
2005-11-17
pubmed:meshHeading
pubmed-meshheading:7806494-Amino Acid Sequence, pubmed-meshheading:7806494-Animals, pubmed-meshheading:7806494-Cell Line, pubmed-meshheading:7806494-Chickens, pubmed-meshheading:7806494-Chromatography, Affinity, pubmed-meshheading:7806494-Chromatography, High Pressure Liquid, pubmed-meshheading:7806494-Crk-Associated Substrate Protein, pubmed-meshheading:7806494-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:7806494-Molecular Sequence Data, pubmed-meshheading:7806494-Oncogene Protein v-crk, pubmed-meshheading:7806494-Phosphoproteins, pubmed-meshheading:7806494-Phosphorylation, pubmed-meshheading:7806494-Proteins, pubmed-meshheading:7806494-Rats, pubmed-meshheading:7806494-Retinoblastoma-Like Protein p130, pubmed-meshheading:7806494-Retroviridae Proteins, Oncogenic, pubmed-meshheading:7806494-Transfection
pubmed:year
1994
pubmed:articleTitle
Characterization, partial purification, and peptide sequencing of p130,the main phosphoprotein associated with v-Crk oncoprotein.
pubmed:affiliation
Molecular Biology Division, Jichi Medical School, Tochigi, Japan.
pubmed:publicationType
Journal Article