Endogenous levels of MKNK2 Isoforms in Normal and Cancer Cell Lines
Disciplines
Biochemistry
Abstract (300 words maximum)
This is part of a Course Based Undergraduate Experience (CURE) for CHEM 3512L. MAP kinase-interacting serine/threonine-protein kinase 2 (MKNK2) has a tumor suppressant long isoform (A) and a pro-oncogenic short isoform (B) which have been established to both be naturally expressed in cells. MKNK2A functions by phosphorylating enzyme p38α, which causes apoptosis and therefore suppresses tumor expression. MKNK2B is not known to phosphorylate p38α and therefore does not lead to apoptosis of tumorous cells and is therefore seen as pro-oncogenic. Pilot studies have been done previously with the MKNK2A and MKNK2B isoforms where both were purified and subjected to a western blot using isoform-specific antibodies. This helped determine whether the antibodies were specific to their respective isoforms. It was found that each isoform-specific antibody was successful in being selective to their respective isoform. The purpose of this project is to establish the natural endogenous level of MKNK2A and MKNK2B in kidney and liver cell lines. The cell lines being tested are HEPG2, derived from liver cells, and BHK, derived from kidney cells, to be analyzed by western blot. Using isoform specific antibodies, the MKNK2A isoform should be visualized as a high intensity band in both cell lines when compared to the MKNK2B isoform due to being upregulated in normal cells. These results of western blotting will give a general idea of if non-overexpressed levels of MKNK2 are able to be significantly measurable in their natural cellular conditions.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Rajnish Singh
Endogenous levels of MKNK2 Isoforms in Normal and Cancer Cell Lines
This is part of a Course Based Undergraduate Experience (CURE) for CHEM 3512L. MAP kinase-interacting serine/threonine-protein kinase 2 (MKNK2) has a tumor suppressant long isoform (A) and a pro-oncogenic short isoform (B) which have been established to both be naturally expressed in cells. MKNK2A functions by phosphorylating enzyme p38α, which causes apoptosis and therefore suppresses tumor expression. MKNK2B is not known to phosphorylate p38α and therefore does not lead to apoptosis of tumorous cells and is therefore seen as pro-oncogenic. Pilot studies have been done previously with the MKNK2A and MKNK2B isoforms where both were purified and subjected to a western blot using isoform-specific antibodies. This helped determine whether the antibodies were specific to their respective isoforms. It was found that each isoform-specific antibody was successful in being selective to their respective isoform. The purpose of this project is to establish the natural endogenous level of MKNK2A and MKNK2B in kidney and liver cell lines. The cell lines being tested are HEPG2, derived from liver cells, and BHK, derived from kidney cells, to be analyzed by western blot. Using isoform specific antibodies, the MKNK2A isoform should be visualized as a high intensity band in both cell lines when compared to the MKNK2B isoform due to being upregulated in normal cells. These results of western blotting will give a general idea of if non-overexpressed levels of MKNK2 are able to be significantly measurable in their natural cellular conditions.