LonP1 chaperone-like activity is ATPase independent and is mediated by its N-domain
The mitochondrial Lon protease is essential for proteostasis through ATP-dependent proteolysis and suppression of protein aggregation through an unknown mechanism. Here we show in three independent aggregation systems that human Lon protease (LonP1) directly interacts with fibrillar aggregates to prevent further aggregation: LonP1 binds amyloid fibrils and inhibits their growth, independently of its protease and ATPase activities. This aggregation inhibition depends on hexamer stability, and even the N-domain hexamer of LonP1 lacking all catalytic domains inhibited aggregation, which localizes its fibril-binding interface. We propose that chaperone deficiencies in LonP1 mutants that are associated with genetic disease, are caused by reduced hexamer stability or increased turnover. Our results clarify the observed dual protease and chaperone function of LonP1 by localizing them to different domains and separating the catalytic activities, thereby facilitating targeting the specific functionalities. Further, we identify the structure of the chaperone substrate to be fibrillar aggregates, suggesting that LonP1 may protect against amyloid fibrils in healthy individuals. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=176 SRC="FIGDIR/small/723147v3_ufig1.gif" ALT="Figure 1"> View larger version (17K): org.highwire.dtl.DTLVardef@1715a1eorg.highwire.dtl.DTLVardef@d382fborg.highwire.dtl.DTLVardef@3c3fa0org.highwire.dtl.DTLVardef@1753f3e_HPS_FORMAT_FIGEXP M_FIG C_FIG SignificanceThe mitochondrial Lon protease has long been proposed to function both as a protease and as a chaperone, though the mechanism of its chaperone activity is debated. Here, we show that human Lon binds to fibrillar protein aggregates and inhibits their elongation, but do not find evidence for chaperoning unfolded chains. Further, our findings challenge the current view that ATPase activity is required for Lon chaperone function. Instead, our results suggest that chaperone deficiency of Lon variants can be explained by variant stability. Our results provide a mechanistic separation of the protease and chaperone-like function LonP1, thereby opening up for targeting one of the functions specifically, and provide new insight into how Lon dysfunction may contribute in multiple ways to age-related and proteostasis-related diseases.