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Purification of Extracellular Trypanosomes, Including African, from Blood by Anion-Exchangers (Diethylaminoethyl-cellulose Columns)
Published on: April 6, 2019
Casein kinase 1 isoform 2 is essential for bloodstream form Trypanosoma brucei
1Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK. m.d.urbaniak@dundee.ac.uk
Abstract:
Induction of RNA interference targeted against casein kinase 1 isoform 2 (TbCK1.2, Tb927.5.800) in bloodstream form Trypanosoma brucei in vitro results in rapid cessation of growth, gross morphological changes, multinucleation and ultimately cell death. A null mutant of the highly homologous casein kinase 1 isoform 1 (Tb927.5.790) in bloodstream form T. brucei displays no growth or morphological phenotype in vitro. A truncated form of TbCK1.2 expressed in Escherichia coli as a GST fusion produces catalytically active recombinant protein, facilitating screening for small molecule inhibitors. These data show that TbCK1.2 is an attractive target for anti-trypanosomal drug discovery.
Insights
Targeting casein kinase 1 isoform 2 (TbCK1.2) in Trypanosoma brucei halts growth and causes cell death, identifying it as a promising target for new anti-parasitic drugs.
Area of Science:
- Parasitology
- Molecular Biology
- Drug Discovery
Background:
- Trypanosoma brucei causes African sleeping sickness.
- Identifying essential genes in T. brucei is crucial for developing treatments.
- Casein kinase 1 (CK1) enzymes are involved in various cellular processes.
Purpose of the Study:
- To investigate the role of casein kinase 1 isoform 2 (TbCK1.2) in T. brucei.
- To assess TbCK1.2 as a potential drug target for trypanosomiasis.
Main Methods:
- RNA interference (RNAi) was used to inhibit TbCK1.2 expression in bloodstream form T. brucei.
- A null mutant for the homologous casein kinase 1 isoform 1 (TbCK1.1) was created.
- Recombinant TbCK1.2 protein was expressed and purified for inhibitor screening.
Main Results:
- TbCK1.2 inhibition led to growth arrest, morphological changes, multinucleation, and cell death.
- TbCK1.1 null mutants showed no observable phenotype in vitro.
- Active recombinant TbCK1.2 protein was successfully produced.
Conclusions:
- TbCK1.2 is essential for the survival of bloodstream form T. brucei.
- TbCK1.2 is a validated and attractive target for anti-trypanosomal drug discovery.
- The recombinant TbCK1.2 protein enables the development of small molecule inhibitors.
