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FAF2 enhances unfoldase activity of mammalian p97-UFD1-NPL4 complex enabling rational design of p97

ABSTRACT: To address how adaptor proteins impact human p97 activity, we here employ in vitro reconstitution and a focused biochemical screen to identify adaptors that enhance unfoldase activity of human p97 in the presence of UFD1-NPL4. Of the adaptors tested, Fas-associated factor 2 (FAF2/UBXD8), a UBX domain adaptor protein, shows the strongest effect on unfolding by p97-UFD1-NPL4 complex. Through biochemical and structural analysis, we define the molecular features that underpin FAF2-mediated stimulation. Based on these insights, we employ computational protein design to engineer de novo mini-proteins that mimic FAF2 function within novel protein scaffolds to potently activate p97. Our work reveals how the unfoldase activity of p97 can be enhanced by additional adaptors, an adaptive feature we believe is important to unfold misfolded proteins, extract proteins from the membrane and tightly bound multiprotein complexes.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Glenn Masson

LAB HEAD: Glenn Masson

PROVIDER: PXD073479 | Pride | 2026-07-28

REPOSITORIES: Pride

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			Action	DRS
	171225_FAF2_ND.raw.zip	Raw
	171225_FAF2_ND_2_IA_final_peptide.csv	Csv
	171225_FAF2_ND_IA_final_peptide.csv	Csv
	211025_P97_UN_FAF2_D_IA_final_peptide.csv	Csv
	221025_P97_UN_FAF2_D_LOW2_IA_final_peptide.csv	Csv

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