Structural basis for the mechanisms of human presequence protease conformational switch and substrate recognition

Year: 2022

Type: article

Abstract: Presequence protease (PreP), a 117 kDa mitochondrial M16C metalloprotease vital for mitochondrial proteostasis, degrades presequence peptides cleaved off from nuclear-encoded proteins and other aggregation-prone peptides, such as amyloid β (Aβ). PreP structures have only been determined in a closed conformation; thus, the mechanisms of substrate binding and selectivity remain elusive. Here, we leverage advanced vitrification techniques to overcome the preferential denaturation of one of two ~55 kDa homologous domains of PreP caused by air-water interface adsorption. Thereby, we elucidate cryoEM structures of three apo-PreP open states along with Aβ- and citrate synthase presequence-bound PreP at 3.3-4.6 Å resolution. Together with integrative biophysical and pharmacological approaches, these structures reveal the key stages of the PreP catalytic cycle and how the binding of substrates or PreP inhibitor drives a rigid body motion of the protein for substrate binding and catalysis. Together, our studies provide key mechanistic insights into M16C metalloproteases for future therapeutic innovations.

Source: Nature Communications

DOI: 10.1038/s41467-022-29322-4

Authors: Wenguang G. Liang, Juwina Wijaya, Hui Wei, Alex J. Noble, Jordan M. Mancl, Swansea Mo, David Lee, John V. Lin King, Man Pan, Chang Liu, Carla M. Koehler, Minglei Zhao, Clinton S. Potter, Bridget Carragher, Sheng Li, Wei-Jen Tang

Institutions: University of Chicago, University of California, Los Angeles, New York Structural Biology Center, University of California San Diego, University of California, San Francisco

Language: English

Topic: Enzyme Structure and Function

Subfield: Materials Chemistry

Field: Materials Science

Domain: Physical Sciences

Open Access status: gold

Estimated APC paid: $5,890

Funders: U.S. Department of Energy, U.S. Department of Health and Human Services, Simons Foundation, National Institutes of Health, National Institute of General Medical Sciences, Basic Energy Sciences

OpenAlex ID: W4226203193