Purification and crystallization of a trimodular complex comprising the type II cohesin–dockerin interaction from the cellulosome of<i>Clostridium thermocellum</i>

Adams, Jarrett; Pal, Gour P.; Yam, Katherine C.; Spencer, Holly L.; Jia, Zongchao; Smith, Steven P.

doi:10.1107/s1744309104025837

Cited by 7 publications

(5 citation statements)

References 25 publications

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“…Indeed, a structural similarity search (33) indicated that the X-module does not share structural similarity with other known X-modules from cellulolytic bacteria, e.g. X-2 (44) or X-60 (3,45). Instead, it exhibits significant similarity with the G5-1 module (p value ϭ 0.04 with 49 equivalent positions and r.m.s.d.…”

Section: Discussionmentioning

confidence: 99%

Atypical Cohesin-Dockerin Complex Responsible for Cell Surface Attachment of Cellulosomal Components

Salama-Alber¹,

Jobby

Chitayat

et al. 2013

Journal of Biological Chemistry

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Background:The type IIIe CohE-XDoc interaction connects cellulosomal components to the cell wall. Results: The dockerin structure in the CohE-XDoc complex exhibits an atypical calcium-binding loop disrupted by a 13-residue insert. Conclusion:The dockerin inserts evolved to serve as novel structural buttresses that support the stalklike X-module conformation. Significance: The type IIIe CohE-XDoc complex underscores dockerin divergence and provides insight into the determinants for cohesin-dockerin specificity.

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Section: Discussionmentioning

confidence: 99%

Atypical Cohesin-Dockerin Complex Responsible for Cell Surface Attachment of Cellulosomal Components

Salama-Alber¹,

Jobby

Chitayat

et al. 2013

Journal of Biological Chemistry

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“…Recently, the crystal structure of the CttA-XDoc complex with ScaE was solved 30 31 , indicating that the insertions serve to reinforce the stalk like structure of the X module. Another form of X module was found to be involved in C. thermocellum type II interactions 57 . These modules are believed to contribute to the solubility, conformational state, structural and thermal stability and spatial flexibility of the cohesin-dockerin pair.…”

Section: Discussionmentioning

confidence: 99%

Complexity of the Ruminococcus flavefaciens FD-1 cellulosome reflects an expansion of family-related protein-protein interactions

Israeli-Ruimy

Bule

Jindou

et al. 2017

Sci Rep

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Protein-protein interactions play a vital role in cellular processes as exemplified by assembly of the intricate multi-enzyme cellulosome complex. Cellulosomes are assembled by selective high-affinity binding of enzyme-borne dockerin modules to repeated cohesin modules of structural proteins termed scaffoldins. Recent sequencing of the fiber-degrading Ruminococcus flavefaciens FD-1 genome revealed a particularly elaborate cellulosome system. In total, 223 dockerin-bearing ORFs potentially involved in cellulosome assembly and a variety of multi-modular scaffoldins were identified, and the dockerins were classified into six major groups. Here, extensive screening employing three complementary medium- to high-throughput platforms was used to characterize the different cohesin-dockerin specificities. The platforms included (i) cellulose-coated microarray assay, (ii) enzyme-linked immunosorbent assay (ELISA) and (iii) in-vivo co-expression and screening in Escherichia coli. The data revealed a collection of unique cohesin-dockerin interactions and support the functional relevance of dockerin classification into groups. In contrast to observations reported previously, a dual-binding mode is involved in cellulosome cell-surface attachment, whereas single-binding interactions operate for cellulosome integration of enzymes. This sui generis cellulosome model enhances our understanding of the mechanisms governing the remarkable ability of R. flavefaciens to degrade carbohydrates in the bovine rumen and provides a basis for constructing efficient nano-machines applied to biological processes.

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“…The crystal structure of the 1:1 type II Coh-XDoc complex was solved by using single-wavelength anomalous diffraction (SAD) data from a single selenomethionyl crystal (30). The atomic model refined at 2.1-Å resolution and the final statistics are summarized in Table 1, which is published as supporting information on the PNAS web site.…”

Section: Resultsmentioning

confidence: 99%

Mechanism of bacterial cell-surface attachment revealed by the structure of cellulosomal type II cohesin-dockerin complex

Adams

Pal

Jia

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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101

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Bacterial cell-surface attachment of macromolecular complexes maintains the microorganism in close proximity to extracellular substrates and allows for optimal uptake of hydrolytic byproducts. The cellulosome is a large multienzyme complex used by many anaerobic bacteria for the efficient degradation of plant cell-wall polysaccharides. The mechanism of cellulosome retention to the bacterial cell surface involves a calcium-mediated protein-protein interaction between the dockerin (Doc) module from the cellulosomal scaffold and a cohesin (Coh) module of cell-surface proteins located within the proteoglycan layer. Here, we report the structure of an ultra-high-affinity (Ka ‫؍‬ 1.44 ؋ 10 10 M ؊1 ) complex between type II Doc, together with its neighboring X module from the cellulosome scaffold of Clostridium thermocellum, and a type II Coh module associated with the bacterial cell surface. Identification of X module-Doc and X module-Coh contacts reveal roles for the X module in Doc stability and enhanced Coh recognition. This extremely tight interaction involves one face of the Coh and both helices of the Doc and comprises significant hydrophobic character and a complementary extensive hydrogen-bond network. This structure represents a unique mechanism for cellsurface attachment in anaerobic bacteria and provides a rationale for discriminating between type I and type II Coh modules.cellulose degradation ͉ cellulosome ͉ protein-protein interaction ͉ calcium A naerobic bacteria rely on secreted hydrolytic enzymes for the breakdown of extracellular polysaccharides into carbon sources, which are readily taken up by the microbe and metabolized. Attachment of these enzymes to the bacterial cell surface, either as independent entities or components of multienzyme complexes, is a key mechanism for the optimal uptake of byproducts and, thus the viability of the microbe, through maintaining its close proximity to extracellular substrates and byproducts. One such complex is the cellulosome, which is responsible for the degradation of crystalline cellulose and associated plant cell-wall polysaccharides (1-6). The cellulosome from Clostridium thermocellum has been the most extensively studied of those identified to date and comprises various cellulases and hemicellulases anchored to a large, multimodular, noncatalytic scaffoldin subunit (CipA). An enzyme-associated calcium (Ca 2ϩ )-binding module termed type I Doc mediates enzyme attachment to the scaffoldin subunit through high affinity noncovalent interactions with its nine highly conserved type I cohesin (Coh) modules yet displays very little preference for particular CipA Coh modules (7,8). The balance of the scaffoldin modular architecture includes a cellulose-binding domain, an X module of unknown function, and a C-terminal type II dockerin (Doc) module. Type II Doc tethers the cellulosome to the proteoglycan layer of the bacterial cell surface through high-affinity interactions with type II Coh modules of the surface-layer homology-containing cell-surface proteins SdbA, O...

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Purification and crystallization of a trimodular complex comprising the type II cohesin–dockerin interaction from the cellulosome ofClostridium thermocellum

Cited by 7 publications

References 25 publications

Atypical Cohesin-Dockerin Complex Responsible for Cell Surface Attachment of Cellulosomal Components

Atypical Cohesin-Dockerin Complex Responsible for Cell Surface Attachment of Cellulosomal Components

Complexity of the Ruminococcus flavefaciens FD-1 cellulosome reflects an expansion of family-related protein-protein interactions

Mechanism of bacterial cell-surface attachment revealed by the structure of cellulosomal type II cohesin-dockerin complex

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