Investigating the isoform specific differences of MKNK2 a/b
Disciplines
Biochemistry, Biophysics, and Structural Biology
Abstract (300 words maximum)
Mitogen-activated protein kinase-interacting kinases (MKNK 1/2s) are important modulators of the MAPK pathways, which control cellular responses such as cell survival, proliferation and oncogenesis for ERK and stress signals for p38. MKNKs are encoded by two genes, presumably via a gene duplication event. Alternative splicing of MKNK2 results in 2 isoforms which can both phosphorylate eIF4E, a key translation initiation factor. The impact of this phosphorylation on translation is still poorly understood. These isoforms vary in their activity, regulation, and subcellular localization. MKNK proteins together can play a role in tumor development, cell migration, cell invasion, and energy metabolism. Understanding the function of MNK would take us one step closer to discovering ways to inhibit it. For this study, we successfully transformed GST-tagged MKNK2 isoforms, long and short, and then produced protein in the presence and absence of Zinc (II) ion. The isoforms were purified using glutathione agarose and then thrombin cleaved to remove the GST-tag. The isoforms were tested for kinase activity and will be tested for zinc content.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Carol Chrestensen
Investigating the isoform specific differences of MKNK2 a/b
Mitogen-activated protein kinase-interacting kinases (MKNK 1/2s) are important modulators of the MAPK pathways, which control cellular responses such as cell survival, proliferation and oncogenesis for ERK and stress signals for p38. MKNKs are encoded by two genes, presumably via a gene duplication event. Alternative splicing of MKNK2 results in 2 isoforms which can both phosphorylate eIF4E, a key translation initiation factor. The impact of this phosphorylation on translation is still poorly understood. These isoforms vary in their activity, regulation, and subcellular localization. MKNK proteins together can play a role in tumor development, cell migration, cell invasion, and energy metabolism. Understanding the function of MNK would take us one step closer to discovering ways to inhibit it. For this study, we successfully transformed GST-tagged MKNK2 isoforms, long and short, and then produced protein in the presence and absence of Zinc (II) ion. The isoforms were purified using glutathione agarose and then thrombin cleaved to remove the GST-tag. The isoforms were tested for kinase activity and will be tested for zinc content.