The Clostridium cellulolyticum Dockerin Displays a Dual Binding Mode for Its Cohesin Partner

Pinheiro, Benedita A.; Proctor, Mark R.; Martinez-Fleites, C.; Prates, José A. M.; Money, Victoria A.; Davies, G.J.; Bayer, Edward A.; Fontes, Carlos M. G. A.; Fierobe, Henri‐Pierre; Gilbert, Harry J.

doi:10.1074/jbc.m801533200

Cited by 76 publications

(133 citation statements)

References 34 publications

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“…ITC experiments were carried out essentially as described previously (11,12), except that the titrations were at 55°C, and proteins were in 50 mM Na-HEPES buffer, pH 7.5, containing 2 mM CaCl 2 . During titration, the dockerin (40 M) was stirred at 300 rpm in the reaction cell, which was injected with 28 successive 10-l aliquots of ligand comprising cohesin (180 M) at 200-s intervals.…”

Section: Isothermal Titration Calorimetry (Itc)mentioning

confidence: 99%

“…Phasing was performed by molecular replacement with the program BALBES (18) using a search model based on the Protein Data Bank structures 2ccl, 1aoh, 2vn6, 1nv8, and 1ixh, mainly related to cohesin and dockerin modules from C. thermocellum and C. cellulolyticum (11,12). Density modification, together with non-crystallographic symmetry averaging, was done with the DM program from the Novel Type I Coh-Doc Complexes from C. thermocellum CCP4 suite (16).…”

Section: Structure Determination and Refinementmentioning

confidence: 99%

“…Dockerins fold into two ␣-helices and EF-hand calcium-binding loop motifs, each corresponding to one of the two duplicated segments (10,12). Thus, the structure of the N-terminal ␣-helix and EF-hand calcium-binding loop can be precisely superimposed over the equivalent structures at the C-terminal end, leading to an internal 2-fold symmetry in the dockerin molecule (11).…”

mentioning

confidence: 99%

“…The genome sequence of C. thermocellum ATCC 27405 encodes 72 polypeptides containing type I dockerin sequences. Alignment of the 72 dockerin sequences at the two ligand binding sites revealed a strong conservation of the amino acids that mediate cohesin recognition (particularly Ser 11 , Thr 12 , and a Lys-Arg motif at positions 18 and 19). Recently, we described the identification of four dockerins, of proteins Cthe_0435 (Cel124A), Cthe_0918, Cthe_0258, and Cthe_0624 (Cel9D-Cel44A), which deviate from the canonical C. thermocellum motifs at least in one of the cohesin binding interfaces (13).…”

mentioning

confidence: 99%

“…Structural studies on type I Coh-Doc complexes of C. thermocellum (10,11) and Clostridium cellulolyticum (12), a mesophilic bacterium that produces a cellulosome analogous to the former microorganism, provided insights into the molecular determinants of protein-protein recognition that mediate the assembly of these protein complexes. Dockerins fold into two ␣-helices and EF-hand calcium-binding loop motifs, each corresponding to one of the two duplicated segments (10,12).…”

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confidence: 99%

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Novel Clostridium thermocellum Type I Cohesin-Dockerin Complexes Reveal a Single Binding Mode

Brás¹,

Alves²,

Carvalho

et al. 2012

Journal of Biological Chemistry

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Section: Isothermal Titration Calorimetry (Itc)mentioning

confidence: 99%

Section: Structure Determination and Refinementmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Novel Clostridium thermocellum Type I Cohesin-Dockerin Complexes Reveal a Single Binding Mode

Brás¹,

Alves²,

Carvalho

et al. 2012

Journal of Biological Chemistry

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A cellulosomal double‐dockerin module from Clostridium thermocellum shows distinct structural and cohesin‐binding features

Chen,

Yang,

Dong

et al. 2024

Protein Science

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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.

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Cohesin diversity revealed by the crystal structure of the anchoring cohesin from Ruminococcus flavefaciens

et al. 2009

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The cellulosome is an intriguing multienzyme complex found in cellulolytic bacteria that plays a key role in the degradation of plant cell-wall polysaccharides. In Ruminococcus flavefaciens, a predominant fiber-degrading bacterium found in ruminants, the cellulosome is anchored to the bacterial cell wall through a relatively short ScaE scaffoldin. Determination of the crystal structure of the lone type-III ScaE cohesin from R. flavefaciens (Rf-CohE) was initiated as a part of a structural effort to define cellulosome assembly. The structure was determined at 1.95 A resolution by single-wavelength anomalous diffraction. This is the first detailed description of a crystal structure for a type-III cohesin, and its features were compared with those of the known type-I and type-II cohesin structures. The Rf-CohE module folds into a nine-stranded beta-sandwich with jellyroll topology, typically observed for cohesins, and includes two beta-flaps in the midst of beta-strands 4 and 8, similar to the type-II cohesin structures. However, the presence in Rf-CohE of an additional 13-residue alpha-helix located between beta-strands 8 and 9 represents a dramatic divergence from other known cohesin structures. The prominent alpha-helix is enveloped by an extensive N-terminal loop, not observed in any other known cohesin, which embraces the helix presumably enhancing its stability. A planar surface at the upper portion of the front face of the molecule, bordered by beta-flap 8, exhibits plausible dimensions and exposed amino acid residues to accommodate the dockerin-binding site.

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The Clostridium cellulolyticum Dockerin Displays a Dual Binding Mode for Its Cohesin Partner

Cited by 76 publications

References 34 publications

Novel Clostridium thermocellum Type I Cohesin-Dockerin Complexes Reveal a Single Binding Mode

Novel Clostridium thermocellum Type I Cohesin-Dockerin Complexes Reveal a Single Binding Mode

A cellulosomal double‐dockerin module from Clostridium thermocellum shows distinct structural and cohesin‐binding features

Cohesin diversity revealed by the crystal structure of the anchoring cohesin from Ruminococcus flavefaciens

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