Cardiac ischemia is a leading cause of death worldwide. The most common and effective treatment is by early reperfusion. However, early reperfusion itself damages the cardiac tissue in a process termed Ischemia reperfusion injury (I/R). Activation of ERK signaling may promote cardioprotection in I/R. ZnT1, a protein that confers resistance to zinc toxicity, was found to interact with Raf1 kinase through its C-terminal domain (CT), leading to downstream activation of ERK in oocytes and C.elegans. In this study, we investigated ZnT1’s ability to activate the MAPK pathway in cultured murine cardiomyocytes (HL1 cells) and its effect during acidic deprivation of oxidative glucose metabolism, an established model for cardiac I/R. Activation of the MAPK pathway was assessed by levels of ERK1/2 phosphorilation, and cellular injury was evaluated by lactate dehydrogenase (LDH) release and staining for proapoptotic caspase activation. Overexpression of ZnT1 markedly increased the basal activation of ERK in response to I/R. Furthermore, overexpressing ZnT1 reduced LDH release and caspase activation following I/R in HL1 cells. This phenomenon was abolished by pretreatment of the cells with the MEK inhibitor PD98059. Knockdown of endogenous ZnT1 reduced phospho-ERK and augmented the I/R induced release of LDH and increased caspase activity. Next we tested the interaction of ZnT1 with Raf1 kinase through its CT, assessing its relevance in this model and in vivo by showing FRET between Raf and ZnT1 in HEK-T293 cells. Furthermore, a truncated form of ZnT1, lacking the CT, failed to activate ERK and did not protect the cells from I/R. In contrast, expression of the CT by itself was sufficient to induce ERK activation and I/R protection. Our findings suggest ZnT1 as a protective candidate protein which augments the activation of the MAPK pathway in the ischemic myocardium through its ability to interact with Raf1 kinase.