Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has historically been viewed only as a glycolytic enzyme and protein; however, increasing recognition as a multi-functional protein, GAPDH serves various roles outside of a its metabolic pathway. Recent studies have demonstrated its involvement in a non-metabolic cellular function, including membrane transport and binding to plasminogen. In the context of intracellular trafficking, GAPDH can interact with members of the cytoskeleton to facilitate regulated transport of vesicles and membrane dynamics in addition to its metabolic functions. The established glycolytic enzyme functions of GAPDH are enhanced as it is now known to bind plasminogen, which is a well-studied precursor to mediating fibrinolysis. This suggests GAPDH serves a role in cellular migration and matrix remodelling. This chapter will address some implications from the idea GAPDH is multi-functional. Through this chapter, we would like to highlight the structural and functional roles on cellular membranes as well as the plasminogen system. As we cover these emerging roles for GAPDH, we also hope to provide some perspectives on the physiological and pathological roles, including potential for future development as a therapeutic target for disorders characterised by aberrant transport and proteolysis.