Iron-sulfur cluster biogenesis is a complex， but highly regulated process that involves de novo cluster formation from iron and sulfide ions on a scaffold protein， and subsequent delivery to final targets via a series of Fe-S cluster-binding carrier proteins. The process of cluster release from the scaffold/carrier for transfer to the target proteins may be mediated by a dedicated Fe-S cluster chaperone system. In human cells， the chaperones include heat shock protein HSPA9 and the J-type chaperone Hsc20. While the role of chaperones has been somewhat clarified in yeast and bacterial systems， many questions remain over their functional roles in cluster delivery and interactions with a variety of human Fe-S cluster proteins. One such protein， Nfu， has recently been recognized as a potential interaction partner of the chaperone complex. Herein， we examined the ability of human Nfu to function as a carrier by interacting with the human chaperone complex. Human Nfu is shown to bind to both chaperone proteins with binding affinities similar to those observed for IscU binding to the homologous HSPA9 and Hsc20， while Nfu can also stimulate the ATPase activity of HSPA9. Additionally， the chaperone complex was able to promote Nfu function by enhancing the second-order rate constants for Fe-S cluster transfer to target proteins and providing directionality in cluster transfer from Nfu by eliminating promiscuous transfer reactions. Together， these data support a hypothesis in which Nfu can serve as an alternative carrier protein for chaperone-mediated cluster release and delivery in Fe-S cluster biogenesis and trafficking.