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:7/28/2014)...   JOLT , a leading technology accelerator, announced today ... portfolio of startups. Next-level tech and boundary-pushing innovation have ... accelerator, and Cohort four is no exception, with six ... its launch in July 2012, JOLT has produced 17 ... at 23 investments, JOLT,s portfolio has an overall mix ...
(Date:7/28/2014)... --  Propeller Health , the FDA-cleared digital health solution ... Brad Towle as Senior Vice President, Business and ... company,s sales growth, customer acquisition and program expansion. ... past role in the development of accountable care organizations, ... time in the industry," said David Van Sickle ...
(Date:7/28/2014)... July 28, 2014  InterMune, Inc. (NASDAQ: ... second quarter 2014 financial results at the close of ... conference call and webcast will be hosted by InterMune ... Interested investors and others may participate in the ... (international), conference ID#21728775. A replay of the webcast and teleconference ...
(Date:7/26/2014)... Albany, NY (PRWEB) July 26, 2014 ... 2020 provides key market data on the Canada ... in millions of US dollars, volume (in units) ... segments - High-End Endoscopy Visualization Systems, Mid-Range Endoscopy ... Browse Full Report at http://www.marketresearchreports.biz/analysis/213254 . , ...
Breaking Biology Technology:JOLT Announces the Next 6 Startups to Join its Summer 2014 Cohort 2JOLT Announces the Next 6 Startups to Join its Summer 2014 Cohort 3Propeller Health Hires Brad Towle as Senior Vice President of Business and Client Development 2InterMune to Release Second Quarter Financial Results on August 6 2Canada Endoscopy Visualization Systems Market Outlook to 2020: MarketResearchReports.Biz 2Canada Endoscopy Visualization Systems Market Outlook to 2020: MarketResearchReports.Biz 3Canada Endoscopy Visualization Systems Market Outlook to 2020: MarketResearchReports.Biz 4Canada Endoscopy Visualization Systems Market Outlook to 2020: MarketResearchReports.Biz 5
... Without effective and proper networking techniques, you may not be ... to other professionals. While people have their own networking style ... , ,You need to do what works best for you ... to learn new ideas about networking from other professionals who ...
... Neurognostics , a home-grown Milwaukee medical device company, finished ... first to take advantage of Wisconsin's new tax credits for ... an investment. , ,Neurognostics oversubscribed on its $1 million angel ... af Neurognostics. As a tech-based research and development company, the ...
... project to match genetic information from Alzheimer's patients with ... ,The two-year project is the first to tap the ... for its Personalized Medicine Research Project, one of the ... of electronic medical records, the project aims to develop ...
Cached Biology Technology:Improve your networking skills by learning from other professionals 2Improve your networking skills by learning from other professionals 3Improve your networking skills by learning from other professionals 4Milwaukee firm among first to use Act 255 tax credits in investor pitch 2Milwaukee firm among first to use Act 255 tax credits in investor pitch 3Milwaukee firm among first to use Act 255 tax credits in investor pitch 4Marshfield study mixes DNA into electronic medical record to screen for Alzheimer's 2Marshfield study mixes DNA into electronic medical record to screen for Alzheimer's 3
(Date:7/28/2014)... He calls himself the bug hunter, but the target ... be found and identified with special methods and instruments. ... State Veterinary Diagnostic Laboratory at Kansas State University, recently ... enterovirus G, which is an important find in the ... cells, but didn,t know what it was," Hause said. ...
(Date:7/28/2014)... July 28, 2014 Research and Markets ... Market in Latin America 2014-2018" report to their ... Biometrics measure and analyze biological data. These systems ... eye retinas, voice patterns, and hand measurements from the ... stored templates. The main purpose of a biometrics system ...
(Date:7/28/2014)... became the first man to reach the South Pole ... an international team of scientists led by Joe McConnell ... air pollution from industrial activities arrived long before. , ... spaced locations around the Antarctic continent, including the South ... reconstruction to date of lead pollution over the Earth,s ...
Breaking Biology News(10 mins):Researcher using next-generation sequencing to rapidly identify pathogens 2Researcher using next-generation sequencing to rapidly identify pathogens 3Biometrics Market in Latin America 2014-2018 2Industrial lead pollution beat explorers to the South Pole by 22 years and persists today 2Industrial lead pollution beat explorers to the South Pole by 22 years and persists today 3
... used as weapons is truly dreadful, but there is plenty ... ancient times. * Bioterrorism, as it is dubbed, ... a safer place against a myriad of old scourges both ... to uncork a nasty cloud of germs. The ...
... oven technology that most people use to heat up leftover ... industry, providing a new way to make thin-film photovoltaic products ... Oregon State University have for the first time developed a ... zinc tin sulfide, a promising solar cell compound that is ...
... many mutations from their fathers as from their mothers, and ... mother,s, researchers have found in the largest study of human ... DNA of around 85,000 Icelanders, also calculates the rate of ... ancestors diverged from nonhuman primates. It is one of two ...
Cached Biology News:Boston subway system to be used to test new sensors for biological agents 2Microwave ovens may help produce lower cost solar energy technology 2Most mutations come from dad 2Most mutations come from dad 3
Ultra-sensitive, non-radioactive detection of Protein G. Measure Protein G levels quickly and easily....
Request Info...
TRIS-EDTA based buffer prepared from RNase-free water suitable for storage and dilution of siRNA, pH 8, store at 4 C ...
Request Info...
Biology Products: