Navigation Links
TKB1 Cells Identify Receptor Tyrosine Kinase Interacting Proteins


In vivo phosphorylation with TKB1 competent cells

David L. Dankort William J. Muller
Institute for Molecular Biology and Biotechnology, McMaster University, Ontario, Canada

GST-fusion proteins expressed in TKB1 bacteria are used to affinity purify cellular proteins, which interact with a specific autophosphorylation site on the ErbB2/Neu receptor tyrosine kinase (RTK). This approach is aptly suited to rapidly identify phosphotyrosine-protein interactions.

When receptor tyrosine kinases (RTKs) are activated, a number of proteins within the cell are rapidly phosphorylated, with the predominate tyrosine-phosphorylated proteins being the RTKs themselves. RTK tyrosine phosphorylation primarily functions to generate binding sites for cytoplasmic- or plasma membrane-associated proteins involved in transducing proliferative or differentiating signals to the nucleus. These signaling proteins each contain modular Src homology 2 (SH2) or protein tyrosine binding/interacting domains (PTB/PID), which enable them to directly interact with specific receptors in a phosphotyrosine-dependent and sequence-specific manner and, therefore, bind to short linear peptide sequences.1,2,3

Extensive mutagenesis of known autophosphorylation sites in the RTK Neu/ErbB-2 has revealed that four (Y1144, Y1201, Y1227, Y1253) of the five autophosphorylation sites are capable of mediating transformation from the receptor.4 Subsequent coimmunoprecipitation analyses from stable cell lines disclose that SHC and GRB2 proteins bind directly to the receptor at tyrosines 1227 and 1144, respectively. 4

Signaling Molecules Identified

Figure 1

To more quickly identify candidate signaling molecules, we developed an in vitro association assay based on the work of Pawson, et al.,5'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. An Epitope Tagging Vector for Gene Expression in Mammalian Cells
2. Versatile Vectors for Ponasterone A- Inducible Control of Gene Expression in Mammalian Cells
3. Cell Proliferation Kit Outperforms the Competition in the Range of Cells Typically Counted
4. XL10-Gold Cells Supplied in New Single-Reaction Format
5. Isolate and Analyze Total RNA from Cells Harvested by Laser Capture Microdissection
6. Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells
7. Simple Isolation of RNA from Tissue and Cultured Cells
8. New BL21-CodonPlus Cells Correct Codon Bias in GC-Rich Genomes
9. Tools for Detecting MSH2 Expression in Chinese Hamster Ovary Cells
10. Low-Toxicity, Lipid-Mediated Transfection of Mammalian Cells
11. Mycoplasma Contamination Reduces the Effect of Lipid-Mediated Transfection of Mammalian Cells
Post Your Comments:
(Date:8/27/2014)... BOSTON , Aug. 27, 2014 Rhythm, ... diseases and genetic deficiencies that result in metabolic disorders, ... on Form S-1 with the U.S. Securities and Exchange ... of shares of its common stock. The number of ... the offering have not yet been determined. ...
(Date:8/27/2014)... 2014 Reportlinker.com announces that a ... its catalogue: Global Biodegradable Packaging ... http://www.reportlinker.com/p02316661/Global-Biodegradable-Packaging-Materials-Market-2014-2018.html About Biodegradable Packaging ... readily decomposed by the action of microorganisms. ... in terms of raw materials, production technology, ...
(Date:8/27/2014)... of nature,s mysteries is how plants survive impact ... the sun,s rays, while using this energy for ... in the plant,s blades quickly dissipate the energy ... quakes. Researchers at DTU Physics have now managed ... algae and bacteria contain light-absorbing proteins which play ...
(Date:8/27/2014)... , Aug. 27, 2014 Reportlinker.com ... available in its catalogue: Global Chelating ... About Chelating Agent A chelating ... covalent bonds with metal ions, thereby forming a ... some metal-ions have on chemical processes, formulations, and ...
Breaking Biology Technology:Rhythm Files Registration Statement for Proposed Initial Public Offering 2Global Biodegradable Packaging Materials Market 2014-2018 2Global Biodegradable Packaging Materials Market 2014-2018 3Global Biodegradable Packaging Materials Market 2014-2018 4Global Biodegradable Packaging Materials Market 2014-2018 5Global Biodegradable Packaging Materials Market 2014-2018 6Global Biodegradable Packaging Materials Market 2014-2018 7DTU researchers film protein quake for the first time 2Global Chelating Agents Market 2014-2018 2Global Chelating Agents Market 2014-2018 3Global Chelating Agents Market 2014-2018 4
... Alfalight , a firm that makes high-power ... has established ties with Chinese distributor Sanetronic Company ... Hong Kong. The company was founded in 1987 as ... has eight offices in China, including Beijing, Shanghai, Shenzhen, ...
... research, nanotechnology, farm waste management and produce storage have emerged ... Transfer Award. , ,Judges for the contest have winnowed the ... The $5,000 grand prize will be presented to the winner ... in Milwaukee. , ,The finalists are: , , GHD ...
... The Joint Finance Committee rejected Governor Jim Doyle's proposal for ... The proposal, a plan to charge consumers for their music, ... several proposals turned down by the budget review panel. , ... on electronic entertainment called by some detractors the "iPod ...
Cached Biology Technology:Five remaining firms vie for Tech Transfer award 2Five remaining firms vie for Tech Transfer award 3Finance Committee rejects extra tax on Internet buys 2Finance Committee rejects extra tax on Internet buys 3
(Date:8/27/2014)... effective doses of the HIV drug maraviroc, a new ... completed before the drug was licensed in 2007, included ... key to removing maraviroc from the body. The current ... protein including nearly half of African-Americans end ... those who lack the protein even when given the ...
(Date:8/27/2014)... neural circuits in the brain of mice, scientists ... The research, led by Howard Hughes Medical Institute ... Technology (MIT), reveals that the connections between the ... about an experience and the part of the ... experience are malleable. , Altering those connections ...
(Date:8/27/2014)... to listen carefully to someone, the first thing we ... is stop moving altogether. This strategy helps us hear ... movements. , This interplay between movement and hearing also ... evidence has long suggested that the brain,s motor cortex, ... gives rise to our conscious perception of sound. , ...
Breaking Biology News(10 mins):Dosage of HIV drug may be ineffective for half of African-Americans 2Researchers change the emotional association of memories 2Researchers change the emotional association of memories 3Researchers change the emotional association of memories 4Stop and listen: Study shows how movement affects hearing 2Stop and listen: Study shows how movement affects hearing 3
... ORLANDO, Fla. Soy food consumption did not ... survivors of breast cancer, according to the results of ... 2011, held April 2-6. Researchers investigated the association ... survivors, using data from a multi-institution collaborative study, the ...
... America (ESA) has selected 37 insect-science symposia for its ... http://entsoc.org/entomology2011 ). The symposia will address important ... Disorder, insect control, invasive species, medical and veterinary entomology, ... BEES AND COLONY COLLAPSE DISORDER Bee ...
... premature aging of immune cells, which may make people more ... new UCSF-led study. The findings indicate that accelerated ... is dependent upon how long someone is depressed, particularly if ... March 2011 by the journal PLoS One and ...
Cached Biology News:Soy isoflavones not a risk for breast cancer survivors 2Entomology 2011 symposia selected 2Entomology 2011 symposia selected 3Study finds link between chronic depression and accelerated immune cell aging 2Study finds link between chronic depression and accelerated immune cell aging 3
... This CLS number is a new product ... number. If showing no availability yet, please ... or contact customer service for assistance. ... Comp Dim: surface area 500 cm 2 ...
... Ideal for cell culture scale-up, these ... 10 7 - 10 9 range ... bottles or spinner flasks. • High mechanical ... leak tested prior to shipping • Certified ...
Mouse Anti-Human Killer cell immunoglobulin-like receptor, two domains, long cytoplasmic tail, 4 (KIR2DL4 Monoclonal Antibody Family: New...
Mouse monoclonal [7B9] to beta II Tubulin ( Abpromise for all tested applications). Antigen: Synthetic peptide, CEEEEGEDEA found at the c-terminus of beta II tubulin. Entrez GeneID: 10383 ...
Biology Products: