Allosteric inhibition of HTRA1 activity by a conformational lock mechanism to treat age-related macular degeneration

Gerhardy, Stefan; Ultsch, Mark; Tang, Wanjian; Green, Evan; Holden, Jeffrey K.; Li, Wei; Estevez, Alberto; Arthur, Chris; Tom, Irene; Rohou, Alexis; Kirchhofer, Daniel

doi:10.1038/s41467-022-32760-9

Allosteric inhibition of HTRA1 activity by a conformational lock mechanism to treat age-related macular degeneration

Journal Article · Mon Sep 05 00:00:00 EDT 2022 · Nature Communications

DOI:https://doi.org/10.1038/s41467-022-32760-9· OSTI ID:1903973

^[1]; Ultsch, Mark ^[2]; ^[3]; Green, Evan ^[2]; Holden, Jeffrey K. ^[3]; Li, Wei ^[3]; Estevez, Alberto ^[2]; Arthur, Chris ^[2]; Tom, Irene ^[4]; ^[2]; ^[3]

Genentech, San Francisco, CA (United States). Dept. of Early Discovery Biochemistry; OSTI
Genentech, San Francisco, CA (United States). Dept. of Structural Biology
Genentech, San Francisco, CA (United States). Dept. of Early Discovery Biochemistry
Genentech, San Francisco, CA (United States). Dept. of OMNI Biomarker Development

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.

View Accepted Manuscript (DOE)

Research Organization:: SLAC National Accelerator Lab., Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL)

Sponsoring Organization:: National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1903973

Journal Information:: Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 13; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
Cryoelectron microsopy
enzyme mechanisms
macular degeneration
proteases
x-ray crystallography

Allosteric inhibition of HTRA1 activity by a conformational lock mechanism to treat age-related macular degeneration

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