Kinase/Phosphatase Substrate Identification Services
IDENTIFY NOVEL KINASE AND PHOSPHATASE SUBSTRATE CONNECTIONS
Take advantage of our “in silico” prediction services to identify sequences of optimal peptide substrates for most protein kinases and our “in vitro” services to discover physiological substrates for kinases and phosphatases.
For more information on our Kinase-Substrate Identification Services please contact our Technical Services representatives toll free at 1-866-KINEXUS or by e-mail at info@kinexus.ca
The vast majority of
the proteins phosphorylated by specific protein kinases in humans and
other species remain unknown despite more than four decades of intense
effort.
As documented in our open-access www.phosphonet.ca and www.kinector.ca knowledgebases, we have identified at least 22 thousand kinase-substrate phosphosite pairs from our literature searches and internal research, but we believe that actual number exceeds 10 million. However, most of this is likely to be collateral phosphorylation and is likely inconsequential.
With the emergence of protein kinases as one of the most promising families of drug targets in the pharmaceutical industry today, it is vital to define the proteins and their most critical phosphosites that are controlled by these important regulatory enzymes.
We expect that many of the genetic mutations that facilitate disease arise from alterations in the amino acids that define the specificity of protein kinases and in the amino acid residues in phosphosites that provide for kinase recognition.
With the sequencing of hundreds of thousands of human genomes that are anticipated in the coming decade, establishing linkages between kinases, phosphosites and gene mutations will provide a major advance in the application of personalized medicine. Towards this end, please check out our www.drugpronet.ca website for some of the bioinformatics tools that we have developed to identify mutations that affect drug binding to over 600 pharmaceutical drugs.
To help realize this potential, Kinexus offers unique and cost effective services that permit the discovery and validation of kinase substrate connections.
The “in silico” prediction services offered by Kinexus have been created using proprietary software that has been developed by Kinexus and our academic partners at the University of British Columbia and Simon Fraser University.
As documented in our open-access www.phosphonet.ca and www.kinector.ca knowledgebases, we have identified at least 22 thousand kinase-substrate phosphosite pairs from our literature searches and internal research, but we believe that actual number exceeds 10 million. However, most of this is likely to be collateral phosphorylation and is likely inconsequential.
With the emergence of protein kinases as one of the most promising families of drug targets in the pharmaceutical industry today, it is vital to define the proteins and their most critical phosphosites that are controlled by these important regulatory enzymes.
We expect that many of the genetic mutations that facilitate disease arise from alterations in the amino acids that define the specificity of protein kinases and in the amino acid residues in phosphosites that provide for kinase recognition.
With the sequencing of hundreds of thousands of human genomes that are anticipated in the coming decade, establishing linkages between kinases, phosphosites and gene mutations will provide a major advance in the application of personalized medicine. Towards this end, please check out our www.drugpronet.ca website for some of the bioinformatics tools that we have developed to identify mutations that affect drug binding to over 600 pharmaceutical drugs.
To help realize this potential, Kinexus offers unique and cost effective services that permit the discovery and validation of kinase substrate connections.
The “in silico” prediction services offered by Kinexus have been created using proprietary software that has been developed by Kinexus and our academic partners at the University of British Columbia and Simon Fraser University.
The algorithm
for kinase substrate prediction uses our databases of the carefully
aligned sequences of 492 human protein kinase catalytic domains and
10,000 known kinase-substrate phosphosite pairs to permit the
elucidation of phosphosite amino acid frequency matrices for any
classical protein kinase for which the catalytic domain sequence is
known.
With this algorithm, Kinexus has been able to predicted the sequences of optimal peptide substrates for most protein kinases.
Conversely, Kinexus can take any known or putative phosphosite sequence and generate a score for each of 482 human protein kinases for their ability to target the sequence for phosphorylation.
Another novel way that we can identify potentially physiological substrates for protein kinases and protein phosphatases is in combination with our high content, KAM-2025 antibody microarray. Kinexus offers this service with over 360 different recombinant human protein kinases and over 40 protein phosphatases to screen endogenous cell and tissue proteins for physiological substrates in high throughput with further filtering of leads with our in silico kinase substrate predictor algorithm. Our Kinase and Phosphatase Substrate Profiling Immunoglobulin Microarray (SPIMA) services are also adaptable with customer supplied kinases, phosphatases and cell/tissue lysates from other eukaryotic species, since many of the phosphosites targeted on this array are highly conserved in evolution, as demonstrated in www.phosphonet.ca.
While the SPIMA method provides for a novel approach for kinase-substrate discovery, when used in combination with Kinexus KAM-2025 antibody microarray analysis of lysates from living cells treated with agents that activate or inhibit these kinases and phosphatases, it becomes possible to establish that these linkages are physiologically relevant in vivo. For example, activation of a protein kinase with a cellular treatment (indicated with increased phosphorylation of an activating phosphosite in the kinase) can be correlated with increased phosphorylation of the putative substrate protein, and the addition of an inhibitor of the kinase can be correlated with a reduction of the phosphorylation of the putative substrate.
With this algorithm, Kinexus has been able to predicted the sequences of optimal peptide substrates for most protein kinases.
Conversely, Kinexus can take any known or putative phosphosite sequence and generate a score for each of 482 human protein kinases for their ability to target the sequence for phosphorylation.
Another novel way that we can identify potentially physiological substrates for protein kinases and protein phosphatases is in combination with our high content, KAM-2025 antibody microarray. Kinexus offers this service with over 360 different recombinant human protein kinases and over 40 protein phosphatases to screen endogenous cell and tissue proteins for physiological substrates in high throughput with further filtering of leads with our in silico kinase substrate predictor algorithm. Our Kinase and Phosphatase Substrate Profiling Immunoglobulin Microarray (SPIMA) services are also adaptable with customer supplied kinases, phosphatases and cell/tissue lysates from other eukaryotic species, since many of the phosphosites targeted on this array are highly conserved in evolution, as demonstrated in www.phosphonet.ca.
While the SPIMA method provides for a novel approach for kinase-substrate discovery, when used in combination with Kinexus KAM-2025 antibody microarray analysis of lysates from living cells treated with agents that activate or inhibit these kinases and phosphatases, it becomes possible to establish that these linkages are physiologically relevant in vivo. For example, activation of a protein kinase with a cellular treatment (indicated with increased phosphorylation of an activating phosphosite in the kinase) can be correlated with increased phosphorylation of the putative substrate protein, and the addition of an inhibitor of the kinase can be correlated with a reduction of the phosphorylation of the putative substrate.