A cellulosomal double-dockerin module from Clostridium thermocellum shows distinct structural and cohesin-binding features.
NMR
X-ray crystallography
binding affinity
protein complex
protein-protein interaction
scaffolding protein
tandem modules
Journal
Protein science : a publication of the Protein Society
ISSN: 1469-896X
Titre abrégé: Protein Sci
Pays: United States
ID NLM: 9211750
Informations de publication
Date de publication:
Apr 2024
Apr 2024
Historique:
revised:
05
02
2024
received:
20
11
2023
accepted:
07
02
2024
medline:
19
3
2024
pubmed:
19
3
2024
entrez:
19
3
2024
Statut:
ppublish
Résumé
Cellulosomes are intricate cellulose-degrading multi-enzymatic complexes produced by anaerobic bacteria, which are valuable for bioenergy development and biotechnology. Cellulosome assembly relies on the selective interaction between cohesin modules in structural scaffolding proteins (scaffoldins) and dockerin modules in enzymes. Although the number of tandem cohesins in the scaffoldins is believed to determine the complexity of the cellulosomes, tandem dockerins also exist, albeit very rare, in some cellulosomal components whose assembly and functional roles are currently unclear. In this study, we characterized the structure and mode of assembly of a tandem bimodular double-dockerin, which is connected to a putative S8 protease in the cellulosome-producing bacterium, Clostridium thermocellum. Crystal and NMR structures of the double-dockerin revealed two typical type I dockerin folds with significant interactions between them. Interaction analysis by isothermal titration calorimetry and NMR titration experiments revealed that the double-dockerin displays a preference for binding to the cell-wall anchoring scaffoldin ScaD through the first dockerin with a canonical dual-binding mode, while the second dockerin module was unable to bind to any of the tested cohesins. Surprisingly, the double-dockerin showed a much higher affinity to a cohesin from the CipC scaffoldin of Clostridium cellulolyticum than to the resident cohesins from C. thermocellum. These results contribute valuable insights into the structure and assembly of the double-dockerin module, and provide the basis for further functional studies on multiple-dockerin modules and cellulosomal proteases, thus highlighting the complexity and diversity of cellulosomal components.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e4937Subventions
Organisme : National Key Research and Development Program of China
ID : 2023YFC3402300
Organisme : National Natural Science Foundation of China
ID : 32070125
Organisme : National Natural Science Foundation of China
ID : 32200030
Organisme : National Natural Science Foundation of China
ID : 32170051
Organisme : National Natural Science Foundation of China
ID : 32171203
Organisme : National Natural Science Foundation of China
ID : 32070028
Organisme : QIBEBT International Cooperation Project
ID : ICP202304
Organisme : State Key Laboratory of Microbial Technology Open Projects Fund
ID : M2022-01
Organisme : Shandong Provincial Natural Science Foundation
ID : ZR2016CB09
Organisme : Fundamental Research Funds for the Central Universities
ID : FRF-DF-20-09
Organisme : Training Program for Young Teaching Backbone Talents
ID : 2302020JXGGRC-005
Organisme : Major Education and Teaching Reform Research Project
ID : JG2021ZD01
Informations de copyright
© 2024 The Protein Society.
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