Title
The Effect of Mutating the PDZ domains within secreted PDZD2: on its insulinotropic action in INSIE cells,Used
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PDZ domains are one of the most common proteinprotein interaction domains in human. PDZ domain containing 2 protein (PDZD2) contains 6 PDZ domains and is highly expressed in pancreatic beta cells. It undergoes cleavage in the endoplasmic reticulum to produce a secreted form PDZD2 (sPDZD2) which retains the last two PDZ domains of PDZD2. sPDZD2 is a candidate beta cell regulatory factor. In current study, two mutated forms of sPDZD2 were generated with alterations in the amino acid sequence within the carboxylatebinding loop of one of the two PDZ domains, either at the PDZ5 or at the PDZ6 domain. Current study revealed that both the PDZ5mutated and PDZ6mutated sPDZD2 proteins failed to exert the insulinotropic effect that the wildtype sPDZD2 had on INS1E cells when added exogenously in the culture medium. Both mutant proteins also failed to rescue the silencing action of siRNA on the insulinotropic effect of endogenous PDZD2 in INS1E cells. These results suggest that intact carboxylatebinding loops of both PDZ5 and PDZ6 domain are crucial for the insulinotropic effect of sPDZD2 exerting on INS1E cells.
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