OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
The insulin receptor kinase (IRK) is composed of two extracellular a-subunits (135 kDa), which contain the insulin-binding site, and two intracellular b-subunits (95 kDa), containing the protein tyrosine kinase domain. After insulin binding, the insulin-IRK complex internalizes into muscle and fat cells. The IRK substrates of IRS-1 and IRS-2 form complexes, through Src homology region 2 (SH2) domains, with docking molecules such as phosphoinositde-3-kinase (PI3K). The recruitment of PI3K in turn activates phosphoinositide-dependent kinases, which serve in the activation of protein kinase B (Akt) and atypical isoforms of protein kinase C (PKC). These activated kinases then phosphorylate downstream effectors, ultimately promoting the translocation of insulin-sensitive glucose transporter subtype 4 (GLUT-4) to the plasma membrane, to operate in the coordination of glucose metabolism.
Family:
Kinase
Sub Family:
RTK
Class:
Protein Tyrosine
Protein Name:
IR
Uniprot Number:
P06213
Protein Aliases:
CD antigen CD220 [Cleaved into: Insulin receptor subunit alpha; Insulin receptor subunit beta]
Gene Name:
INSR
Gene ID:
3643
Gene Aliases:
CD220
Accession Number:
NM_000208
Organism:
Human
Construct Details:
1005-1310 recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometric
Measured Response:
Scintillation
Testing Information
Procedure Summary:
IR (h) is incubated with 50 mM Tris pH 7.5, 0.1 mM EGTA, 0.1 mM Na3VO4, 0.1% 6-mercaptoethanol, 250 µM KKSRGDYMTMQIG, 10 mM MnCl2, 10 mM MgAcetate and [gamma-33P]-ATP (specific activity and concentration as required). The reaction is initiated by the addition of the Mg/ATP mix. After incubation for 40 minutes at room temperature, the reaction is stopped by the addition of phosphoric acid to a concentration of 0.5%. An aliquot of the reaction is then spotted onto a filter and washed four times for 4 minutes in 0.425% phosphoric acid and once in methanol prior to drying and scintillation counting.
Substrate:
250 uM KKSRGDYMTMQIG
Tracer:
33P
ATP Concentration:
[ATP]= 200µM (Km= 447µM )
Incubation:
40 min at Room temperature
Control Inhibitor:
Staurosporine
Test Sample Requirements:
Please provide compounds dissolved in 100% DMSO (PBS or HBSS prohibited).
Stock solution should be ≥50× the final assay concentration (a 10 mM stock is also acceptable).
Required volume:
– <60 kinase assays: 120 µL of 50× stock
– ≥60 assays: 420 µL of 50× stock
If you supply 10 mM stocks or powder (pre-weighed), we will apply the appropriate conversion.
For batches of ~50 compounds supplied as solids, the TAT will be extended.
Minimum Order Quantity:
2
Turnaround Time
Standard:
6 Business Days
Service Scheduling Note:
Assay is run on a weekly schedule beginning on Friday. Compounds and purchase order must be received on or before 13:00 CET Thursday for the order to be included on the service run that begins on Friday of the same week.
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Metabolic Diseases
Adverse / Beneficial Action:
The insulin receptor kinase may be effective in improving glycemic control in diabetics.
Additional Information
Brand:
LeadHunter
Testing Location:
KP - France - Celle L'Evescault
RELATED SOLUTIONS
IR (INSR) Human RTK Kinase Enzymatic Radiometric [10 uM ATP] KinaseProfiler LeadHunter Assay - FR
Item: 14-466KP10
IR (INSR) (Activated) Human RTK Kinase Enzymatic Radiometric [Km ATP] KinaseProfiler LeadHunter Assay - FR
Item: 14-803KP
IR (INSR) (Activated) Human RTK Kinase Enzymatic Radiometric [10 uM ATP] KinaseProfiler LeadHunter Assay - FR
Item: 14-803KP10
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