Cohesin–dockerin interaction in cellulosome assembly: a single Asp‐to‐Asn mutation disrupts high‐affinity cohesin–dockerin binding
- 26 July 2004
- journal article
- Published by Wiley in FEBS Letters
- Vol. 572 (1-3) , 195-200
- https://doi.org/10.1016/j.febslet.2004.07.040
Abstract
The cohesive cellulosome complex is sustained by the high-affinity cohesin–dockerin interaction. In previous work [J. Biol. Chem. 276 (2001) 9883], we demonstrated that a single Thr-to-Leu replacement in the Clostridium thermocellum dockerin component differentiates between non-recognition and high-affinity recognition by the interspecies rival cohesin from C. cellulolyticum. In this report, we show that a single Asp-to-Asn substitution on the cohesin counterpart also disrupts normal recognition of the dockerin. The Asp34 carboxyl group of the cohesin appears to play a central role in the resultant hydrogen-bonding network as an acceptor of two crucial hydrogen bonds from Ser45 of the dockerin domain. The results underscore the fragile nature of the intermolecular contact interactions that maintain this very high-affinity protein–protein interactionKeywords
This publication has 27 references indexed in Scilit:
- Effect of Rational Mutagenesis of Selected Cohesin Residues on the High-Affinity Cohesin-Dockerin InteractionPublished by American Chemical Society (ACS) ,2004
- Mapping by Site-Directed Mutagenesis of the Region Responsible for Cohesin−Dockerin Interaction on the Surface of the Seventh Cohesin Domain of Clostridium thermocellum CipABiochemistry, 2002
- Cohesin-Dockerin Interaction in Cellulosome AssemblyJournal of Biological Chemistry, 2001
- Solution Structure of a Type I Dockerin Domain, a Novel Prokaryotic, Extracellular Calcium-binding Domain11Edited by P. E. WrightJournal of Molecular Biology, 2001
- Crystal structure of a cohesin module from Clostridium cellulolyticum: implications for dockerin recognitionJournal of Molecular Biology, 2000
- The atomic structure of protein-protein recognition sites 1 1Edited by A. R. FershtJournal of Molecular Biology, 1999
- Anatomy of hot spots in protein interfacesJournal of Molecular Biology, 1998
- The crystal structure of a type I cohesin domain at 1.7 Å resolutionJournal of Molecular Biology, 1997
- Expression, purification and subunit‐binding properties of cohesins 2 and 3 of the Clostridium thermocellum cellulosomeFEBS Letters, 1995
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976