Insight into the autoproteolysis mechanism of the RsgI9 anti-σ factor from Clostridium thermocellum.
Clostridium thermocellum
RsgI
anti‐sigma factor
autoproteolysis
cellulosome
conserved RsgI extracellular domain
periplasmic domain
Journal
Proteins
ISSN: 1097-0134
Titre abrégé: Proteins
Pays: United States
ID NLM: 8700181
Informations de publication
Date de publication:
10 Apr 2024
10 Apr 2024
Historique:
revised:
19
03
2024
received:
06
11
2023
accepted:
26
03
2024
medline:
10
4
2024
pubmed:
10
4
2024
entrez:
10
4
2024
Statut:
aheadofprint
Résumé
Clostridium thermocellum is a potential microbial platform to convert abundant plant biomass to biofuels and other renewable chemicals. It efficiently degrades lignocellulosic biomass using a surface displayed cellulosome, a megadalton sized multienzyme containing complex. The enzymatic composition and architecture of the cellulosome is controlled by several transmembrane biomass-sensing RsgI-type anti-σ factors. Recent studies suggest that these factors transduce signals from the cell surface via a conserved RsgI extracellular (CRE) domain (also called a periplasmic domain) that undergoes autoproteolysis through an incompletely understood mechanism. Here we report the structure of the autoproteolyzed CRE domain from the C. thermocellum RsgI9 anti-σ factor, revealing that the cleaved fragments forming this domain associate to form a stable α/β/α sandwich fold. Based on AlphaFold2 modeling, molecular dynamics simulations, and tandem mass spectrometry, we propose that a conserved Asn-Pro bond in RsgI9 autoproteolyzes via a succinimide intermediate whose formation is promoted by a conserved hydrogen bond network holding the scissile peptide bond in a strained conformation. As other RsgI anti-σ factors share sequence homology to RsgI9, they likely autoproteolyze through a similar mechanism.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NIH HHS
ID : S10OD025073
Pays : United States
Organisme : NIH HHS
ID : S10OD016336
Pays : United States
Organisme : NIH HHS
ID : R01-AI052217
Pays : United States
Organisme : NIDCR NIH HHS
ID : T90-DE030860
Pays : United States
Informations de copyright
© 2024 Wiley Periodicals LLC.
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