Ten catalytic snapshots of rhomboid intramembrane proteolysis from gate opening to peptide release.
Journal
Nature structural & molecular biology
ISSN: 1545-9985
Titre abrégé: Nat Struct Mol Biol
Pays: United States
ID NLM: 101186374
Informations de publication
Date de publication:
10 2019
10 2019
Historique:
received:
26
03
2019
accepted:
09
08
2019
pubmed:
2
10
2019
medline:
12
2
2020
entrez:
2
10
2019
Statut:
ppublish
Résumé
Protein cleavage inside the cell membrane triggers various pathophysiological signaling pathways, but the mechanism of catalysis is poorly understood. We solved ten structures of the Escherichia coli rhomboid protease in a bicelle membrane undergoing time-resolved steps that encompass the entire proteolytic reaction on a transmembrane substrate and an aldehyde inhibitor. Extensive gate opening accompanied substrate, but not inhibitor, binding, revealing that substrates and inhibitors take different paths to the active site. Catalysis unexpectedly commenced with, and was guided through subsequent catalytic steps by, motions of an extracellular loop, with local contributions from active site residues. We even captured the elusive tetrahedral intermediate that is uncleaved but covalently attached to the catalytic serine, about which the substrate was forced to bend dramatically. This unexpectedly stable intermediate indicates rhomboid catalysis uses an unprecedented reaction coordinate that may involve mechanically stressing the peptide bond, and could be selectively targeted by inhibitors.
Identifiants
pubmed: 31570873
doi: 10.1038/s41594-019-0296-9
pii: 10.1038/s41594-019-0296-9
pmc: PMC6858540
mid: NIHMS1537267
doi:
Substances chimiques
DNA-Binding Proteins
0
Escherichia coli Proteins
0
GlpG protein, E coli
0
Membrane Proteins
0
Peptides
0
Endopeptidases
EC 3.4.-
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
910-918Subventions
Organisme : NIGMS NIH HHS
ID : P41 GM103485
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI066025
Pays : United States
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