Allosteric inhibition of HTRA1 activity by a conformational lock mechanism to treat age-related macular degeneration.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
05 09 2022
05 09 2022
Historique:
received:
03
02
2022
accepted:
16
08
2022
entrez:
6
9
2022
pubmed:
7
9
2022
medline:
9
9
2022
Statut:
epublish
Résumé
The trimeric serine protease HTRA1 is a genetic risk factor associated with geographic atrophy (GA), a currently untreatable form of age-related macular degeneration. Here, we describe the allosteric inhibition mechanism of HTRA1 by a clinical Fab fragment, currently being evaluated for GA treatment. Using cryo-EM, X-ray crystallography and biochemical assays we identify the exposed LoopA of HTRA1 as the sole Fab epitope, which is approximately 30 Å away from the active site. The cryo-EM structure of the HTRA1:Fab complex in combination with molecular dynamics simulations revealed that Fab binding to LoopA locks HTRA1 in a non-competent conformational state, incapable of supporting catalysis. Moreover, grafting the HTRA1-LoopA epitope onto HTRA2 and HTRA3 transferred the allosteric inhibition mechanism. This suggests a conserved conformational lock mechanism across the HTRA family and a critical role of LoopA for catalysis, which was supported by the reduced activity of HTRA1-3 upon LoopA deletion or perturbation. This study reveals the long-range inhibition mechanism of the clinical Fab and identifies an essential function of the exposed LoopA for activity of HTRA family proteases.
Identifiants
pubmed: 36064790
doi: 10.1038/s41467-022-32760-9
pii: 10.1038/s41467-022-32760-9
pmc: PMC9445180
doi:
Substances chimiques
Epitopes
0
Immunoglobulin Fab Fragments
0
HTRA3 protein, human
EC 3.4.21.-
High-Temperature Requirement A Serine Peptidase 1
EC 3.4.21.-
HTRA1 protein, human
EC 3.4.21.-
Serine Endopeptidases
EC 3.4.21.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
5222Subventions
Organisme : NIGMS NIH HHS
ID : P30 GM133894
Pays : United States
Informations de copyright
© 2022. The Author(s).
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