Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
15
pubmed:dateCreated
2004-4-8
pubmed:abstractText
Constitutive overexpression of nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) is a key oncogenic event in anaplastic large-cell lymphomas with the characteristic chromosomal aberration t(2;5)(p23;q35). Proteins that interact with ALK tyrosine kinase play important roles in mediating downstream cellular signals, and are potential targets for novel therapies. Using a functional proteomic approach, we determined the identity of proteins that interact with the ALK tyrosine kinase by co-immunoprecipitation with anti-ALK antibody, followed by electrospray ionization and tandem mass spectrometry (MS/MS). A total of 46 proteins were identified as unique to the ALK immunocomplex using monoclonal and polyclonal antibodies, while 11 proteins were identified in the NPM immunocomplex. Previously reported proteins in the ALK signal pathway were identified including PI3-K, Jak2, Jak3, Stat3, Grb2, IRS, and PLCgamma1. More importantly, many proteins previously not recognized to be associated with NPM-ALK, but with potential NPM-ALK interacting protein domains, were identified. These include adaptor molecules (SOCS, Rho-GTPase activating protein, RAB35), kinases (MEK kinase 1 and 4, PKC, MLCK, cyclin G-associated kinase, EphA1, JNK kinase, MAP kinase 1), phosphatases (meprin, PTPK, protein phosphatase 2 subunit), and heat shock proteins (Hsp60 precursor). Proteins identified by MS were confirmed by Western blotting and reciprocal immunoprecipitation. This study demonstrates the utility of antibody immunoprecipitation and subsequent peptide identification by tandem mass spectrometry for the elucidation of ALK-binding proteins, and its potential signal transduction pathways.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
0950-9232
pubmed:author
pubmed:issnType
Print
pubmed:day
8
pubmed:volume
23
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
2617-29
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:14968112-Amino Acid Sequence, pubmed-meshheading:14968112-Antibodies, Monoclonal, pubmed-meshheading:14968112-Blotting, Western, pubmed-meshheading:14968112-Cell Line, Tumor, pubmed-meshheading:14968112-Chromosome Aberrations, pubmed-meshheading:14968112-Databases as Topic, pubmed-meshheading:14968112-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:14968112-Humans, pubmed-meshheading:14968112-Mass Spectrometry, pubmed-meshheading:14968112-Molecular Sequence Data, pubmed-meshheading:14968112-Peptides, pubmed-meshheading:14968112-Precipitin Tests, pubmed-meshheading:14968112-Protein Binding, pubmed-meshheading:14968112-Protein-Tyrosine Kinases, pubmed-meshheading:14968112-Proteomics, pubmed-meshheading:14968112-Sequence Homology, Amino Acid, pubmed-meshheading:14968112-Signal Transduction, pubmed-meshheading:14968112-Spectrometry, Mass, Electrospray Ionization, pubmed-meshheading:14968112-Thermodynamics, pubmed-meshheading:14968112-Trypsin
pubmed:year
2004
pubmed:articleTitle
Identification of NPM-ALK interacting proteins by tandem mass spectrometry.
pubmed:affiliation
ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT 84108, USA.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't