Biological Information

Background Information:

The lck protooncogene is a member of the src-related gene family of non-receptor tyrosine kinases. It is uniquely expressed only in T-lymphocytes and NK-cells. The p56lck kinase (LCK) is a 56 KD polypeptide that is associated with a cytoplasmic face of the plasma membrane. It functions as a critical signalling model in IL-2 signal transduction leading to T-lymphocyte activation. Specifically, it has been shown that a stable complex exists between IL-2 receptors and p56lck and that treatment of T-lymphocytes with IL-2 promotes p56lck kinase activity and rapid phosphorylation of cellular substrates. In addition, p56lck has been found to function as a signal transducer for antigen activated CD4 and CG8 T-cell receptors and plays a role in thymic maturation of certain T-cells. The CD45 antigen, a receptor/protein tyrosine phosphatase of T-cells, also interacts with p56lck.

Family:

Kinase

Sub Family:

TK

Class:

Protein Tyrosine

Protein Name:

Lck

Uniprot Number:

P06239

Protein Aliases:

Leukocyte C-terminal Src kinase|LSK|Lymphocyte cell-specific protein-tyrosine kinase|Protein YT16|Proto-oncogene Lck|T cell-specific protein-tyrosine kinase|p56-LCK

Gene Name:

LCK

Gene ID:

3932

Accession Number:

NM_005356

Organism:

Human

Construct Details:

Full Length, Recombinant

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Enzymatic

Functional Mode:

Antagonist

Detection Method:

Radiometry

Measured Response:

Scintillation

Testing Information

Procedure Summary:

Human recombinant protein kinase LCK expressed in insect cells is used. Test compound and/or vehicle is preincubated with 0.4 μg/ml enzyme‡ in modified HEPES buffer pH 7.4 for 15 minutes at 37°C. The reaction is initiated by addition of 0.2 mg/ml poly(Glu:Tyr), 10 μM ATP and 0.25 μCi [γ32P]ATP for another 30 minute incubation period and terminated by further addition of 3% H3PO4. An aliquot is removed and counted to determine the amount of [32P]Poly(Glu:Tyr) formed. Compounds are screened at 10 μM.

Substrate:

200 µg/ml Poly(Glu:Tyr)

ATP Concentration:

10 uM

Incubation:

30 min at 37°C

Control Inhibitor:

TC-S 7003

Criteria For Significance:

≥ 50% of max stimulation or inhibition

Test Concentration / Dose:

Suggested concentration for initial testing is 10 uM

Test Sample Requirements:

Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).

Minimum Order Quantity:

2

Turnaround Time

Standard:

15 Business Days

Reference Compound Data

PubChem ID

122130844

Name

Staurosporine

Measurement

0.054 μM - IC50

PubChem ID

11785878

Name

6-(2,6-Dimethylphenyl)-2-[[4-(4-methyl-1-pyrimido[5',4':5,6]

Measurement

0.064 μM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Adverse / Beneficial Action:

Inhibition of LCK may result in immune suppression and apoptosis.

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

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