biostudies-literatureUnknownLin JNIGMS NIH HHSThe hexameric AAA+ disaggregase, Hsp104, collaborates with Hsp70 and Hsp40 via its autoregulatory middle domain (MD) to solubilize aggregated protein conformers. However, how ATP- or ADP-specific MD configurations regulate Hsp104 hexamers remains poorly understood. Here, we define an ATP-specific network of interprotomer contacts between nucleotide-binding domain 1 (NBD1) and MD helix L1, which tunes Hsp70 collaboration. Manipulating this network can: (a) reduce Hsp70 collaboration without enhancing activity; (b) generate Hsp104 hypomorphs that collaborate selectively with class B Hsp40s; (c) produce Hsp70-independent potentiated variants; or (d) create species barriers between Hsp104 and Hsp70. Conversely, ADP-specific intraprotomer contacts between MD helix L2 and NBD1 restrict activity,bioRxiv : the preprint server for biology2024.04.26.591398https://www.ebi.ac.uk/biostudies/studies/S-EPMC11071516biostudies-literatureDesign principles to tailor Hsp104 therapeutics.PMC11071516T32 GM008076R01 GM099836R01 GM110001Rizo ANGates SNGambogi CWChuang EYokom ALLin JSouthworth DRShorter JCarman PJKendsersky NMfalseDesign principles to tailor Hsp104 therapeutics.The hexameric AAA+ disaggregase, Hsp104, collaborates with Hsp70 and Hsp40 via its autoregulatory middle domain (MD) to solubilize aggregated protein conformers. However, how ATP- or ADP-specific MD configurations regulate Hsp104 hexamers remains poorly understood. Here, we define an ATP-specific network of interprotomer contacts between nucleotide-binding domain 1 (NBD1) and MD helix L1, which tunes Hsp70 collaboration. Manipulating this network can: (a) reduce Hsp70 collaboration without enhancing activity; (b) generate Hsp104 hypomorphs that collaborate selectively with class B Hsp40s; (c) produce Hsp70-independent potentiated variants; or (d) create species barriers between Hsp104 and Hsp70. Conversely, ADP-specific intraprotomer contacts between MD helix L2 and NBD1 restrict activity,2024-01-01T00:00:00Z2024 Apr2026-04-15T03:26:04.994Z2025-08-14T03:05:50.954ZS-EPMC110715163871216810.1101/2024.04.26.591398