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Updated: Mar 22, 2026
Identification of Kinase-substrate Pairs Using High Throughput Screening
Published on: August 29, 2015
Discovery of a Highly Selective STK16 Kinase Inhibitor
Feiyang Liu1,2, Jinhua Wang3, Xingxing Yang1
1High Magnetic Field Laboratory, Chinese Academy of Sciences , 350 Shushanhu Road, P.O. Box 1110, Hefei, Anhui 230031, People's Republic of China.
Abstract:
STK16, a serine/threonine protein kinase, is ubiquitously expressed and is conserved among all eukaryotes. STK16 has been implicated to function in a variety of cellular processes such as VEGF and cargo secretion, but the pathways through which these effects are mediated remain to be elucidated. Through screening of our focused library of kinase inhibitors, we discovered a highly selective ATP competitive inhibitor, STK16-IN-1, which exhibits potent inhibitory activity against STK16 kinase (IC50: 0.295 μM) with excellent selectivity across the kinome as assessed using the KinomeScan profiling assay (S score (1) = 0.0). In MCF-7 cells, treatment with STK16-IN-1 results in a reduction in cell number and accumulation of binucleated cells, which can be recapitulated by RNAi knockdown of STK16. Co-treatment of STK16-IN-1 with chemotherapeutics such as cisplatin, doxorubicin, colchicine, and paclitaxel results in a slight potentiation of the antiproliferative effects of the chemotherapeutics. STK16-IN-1 provides a useful tool compound for further elucidating the biological functions of STK16.
Insights
A novel inhibitor, STK16-IN-1, selectively targets the STK16 protein kinase. This compound reduces cancer cell proliferation and enhances chemotherapy effects, offering a new tool for biological research.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Serine/threonine protein kinase 16 (STK16) is conserved in eukaryotes and involved in cellular processes like secretion.
- The precise mechanisms of STK16's function remain largely unknown.
- Targeted inhibition is crucial for dissecting STK16's biological roles.
Purpose of the Study:
- To identify and characterize a selective inhibitor of STK16.
- To investigate the cellular effects of STK16 inhibition.
- To evaluate the potential of STK16 inhibition in combination cancer therapy.
Main Methods:
- Screening of a focused kinase inhibitor library to discover STK16 inhibitors.
- Biochemical assays to determine inhibitor potency (IC50) and kinome selectivity.
- Cell-based assays (MCF-7 cells) using the inhibitor and RNAi for STK16 knockdown.
- Combination studies with standard chemotherapeutic agents.
Main Results:
- Discovery of STK16-IN-1, a potent and highly selective ATP-competitive inhibitor of STK16 (IC50 = 0.295 μM, S score = 0.0).
- STK16-IN-1 treatment in MCF-7 cells led to decreased cell number and binucleated cell accumulation, mirroring STK16 RNAi effects.
- Co-treatment with STK16-IN-1 slightly potentiated the antiproliferative activity of cisplatin, doxorubicin, colchicine, and paclitaxel.
Conclusions:
- STK16-IN-1 is a valuable chemical probe for studying STK16 function.
- STK16 plays a role in cell proliferation and cytokinesis.
- Inhibition of STK16 may offer a strategy to enhance the efficacy of certain chemotherapies.
