Activity of MKNK2 Isoforms Studied by Phosphorylation with p38a-MAPK
Disciplines
Medicine and Health Sciences
Abstract (300 words maximum)
Mnk2 is a protein kinase that is known to phosphorylate p38a-MAPK. Two isoforms of MNKN2 are known. Isoform A has been studied in detail, however, Isoform B, which is a splice variant and has a shorter sequence and is understudied. Both isoforms are differentially expressed in cancer cells with isoform A expressed much less than at normal levels, and isoform B being expressed at higher levels in cancer cells. The MKNK2 isoform A contains a MAPK binding site which gives the possibility of a binding site for p38a-MAPK. Whereas, the MNKN2 isoform B has been reported to have multiple tyrosine and threonine residues, showing potential to be phosphorylated as well. The purpose of this course based undergraduate research experience is to analyze both isoforms of MKNK2 to see which specific one is phosphorylated by p38-MAPK in order to better understand their function. Preliminary western blot experiment has been performed to ensure the specificity of antibodies for each isoform. A kinase assay coupled with western blotting will be done to study further the phosphorylation of both isoforms MKNK2 by p38a-MAPK. The in vitro kinase assay will use p38 as the enzyme and the purified, his-tagged isoforms as the substrates. The resulting phosphorylation should change the mass of the MKNK2 isoforms, showing the isoforms to be under 80 kD while the isoforms paired with p38a should be above 80 kD on the western blot.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Rajnish Singh
Activity of MKNK2 Isoforms Studied by Phosphorylation with p38a-MAPK
Mnk2 is a protein kinase that is known to phosphorylate p38a-MAPK. Two isoforms of MNKN2 are known. Isoform A has been studied in detail, however, Isoform B, which is a splice variant and has a shorter sequence and is understudied. Both isoforms are differentially expressed in cancer cells with isoform A expressed much less than at normal levels, and isoform B being expressed at higher levels in cancer cells. The MKNK2 isoform A contains a MAPK binding site which gives the possibility of a binding site for p38a-MAPK. Whereas, the MNKN2 isoform B has been reported to have multiple tyrosine and threonine residues, showing potential to be phosphorylated as well. The purpose of this course based undergraduate research experience is to analyze both isoforms of MKNK2 to see which specific one is phosphorylated by p38-MAPK in order to better understand their function. Preliminary western blot experiment has been performed to ensure the specificity of antibodies for each isoform. A kinase assay coupled with western blotting will be done to study further the phosphorylation of both isoforms MKNK2 by p38a-MAPK. The in vitro kinase assay will use p38 as the enzyme and the purified, his-tagged isoforms as the substrates. The resulting phosphorylation should change the mass of the MKNK2 isoforms, showing the isoforms to be under 80 kD while the isoforms paired with p38a should be above 80 kD on the western blot.