| DOI | Resolve DOI: https://doi.org/10.1124/mol.113.089847 |
|---|
| Author | Search for: Perron, M.D.; Search for: Chowdhury, S.; Search for: Aubry, I.; Search for: Purisima, E.1; Search for: Tremblay, M.L.; Search for: Uri Saragovi, H. |
|---|
| Affiliation | - National Research Council of Canada. NRC Biotechnology Research Institute
|
|---|
| Format | Text, Article |
|---|
| Subject | CD45 antigen; compound 211; interleukin 2; mitogen activated protein kinase; protein kinase Lck; protein kinase ZAP 70; protein tyrosine phosphatase inhibitor; T lymphocyte receptor; unclassified drug; animal cell; animal experiment; animal model; assay; binding site; cell activity; computer model; cytokine production; delayed hypersensitivity; drug dose escalation; drug potency; drug selectivity; female; IC 50; immunosuppressive treatment; in vivo study; inflammation; lymphocytic infiltration; molecular docking; mouse; neutropenia; protein dephosphorylation; signal transduction; single drug dose; site directed mutagenesis; spleen cell; T lymphocyte; Allosteric Regulation; Allosteric Site; Antigens, CD45; Cells, Cultured; Enzyme Activation; Female; Hypersensitivity, Delayed; Immunologic Factors; Immunosuppressive Agents; Inflammation; Interleukin-2; Lymphocyte Specific Protein Tyrosine Kinase p56(lck); Mice; Mice, Inbred C57BL; Mitogen-Activated Protein Kinases; Molecular Docking Simulation; Mutagenesis, Site-Directed; Naphthoquinones; Phosphorylation; Phosphotyrosine; Receptors, Antigen, T-Cell; Signal Transduction; Structure-Activity Relationship; ZAP-70 Protein-Tyrosine Kinase |
|---|
| Abstract | CD45 is a receptor-like member of the protein tyrosine phosphatase (PTP) family. We screened in silico for small molecules binding at a predicted allosteric pocket unique to the CD45 intracellular domain, and validated inhibitors by in vitro phosphatase assays. Compound 211 exhibited a CD45 IC 50 value of 200 nM and had >.100-fold selectivity over six related PTPs. The relevance of the allosteric pocket was verified through site-directed mutagenesis. Compound 211 has a noncompetitive mechanism of action, and it is extremely effective at preventing dephosphorylation of substrate Lck phosphotyrosine (pY)-505 versus preventing dephosphorylation of Lck pY-393. In cultured primary T cells, compound 211 prevents T-cell receptor-mediated activation of Lck, Zap-70, and mitogen-activated protein kinase, and interleukin-2 production. In a delayed-type hypersensitivity reaction in vivo, compound 211 abolished inflammation. This work demonstrates a novel approach to develop effective allosteric inhibitors that can be expanded to target the corresponding allosteric domains of other receptor PTPs. |
|---|
| Publication date | 2014 |
|---|
| In | |
|---|
| Language | English |
|---|
| Peer reviewed | Yes |
|---|
| NPARC number | 21272140 |
|---|
| Export citation | Export as RIS |
|---|
| Report a correction | Report a correction (opens in a new tab) |
|---|
| Record identifier | 2cd1e146-72dc-458b-ade6-562045109eda |
|---|
| Record created | 2014-07-23 |
|---|
| Record modified | 2020-04-22 |
|---|
