Crucial roles of single residues in binding affinity, specificity, and promiscuity in the cellulosomal cohesin-dockerin interface

Michal Slutzki, Dan Reshef, Yoav Barak, Rachel Haimovitz, Shahar Rotem-Bamberger, Raphael Lamed, Edward A. Bayer, Ora Schueler-Furman*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Interactions between cohesin and dockerin modules play a crucial role in the assembly of multienzyme cellulosome complexes. Although intraspecies cohesin and dockerin modules bind in general with high affinity but indiscriminately, crossspecies binding is rare. Here, we combined ELISA-based experiments with Rosetta-based computational design to evaluate the contribution of distinct residues at the Clostridium thermocellum cohesin-dockerin interface to binding affinity, specificity, and promiscuity. We found that single mutations can show distinct and significant effects on binding affinity and specificity. In particular, mutations at cohesin position Asn37 show dramatic variability in their effect on dockerin binding affinity and specificity: the N37A mutant binds promiscuously both to cognate (C. thermocellum) as well as to non-cognate Clostridium cellulolyticum dockerin. N37L in turn switches binding specificity: compared with the wild-type C. thermocellum cohesin, this mutant shows significantly increased preference for C. cellulolyticum dockerin combined with strongly reduced binding to its cognate C. thermocellum dockerin. The observation that a single mutation can overcome the naturally observed specificity barrier provides insights into the evolutionary dynamics of this system that allows rapid modulation of binding specificity within a high affinity background.

Original languageEnglish
Pages (from-to)13654-13666
Number of pages13
JournalJournal of Biological Chemistry
Issue number22
StatePublished - 29 May 2015


FundersFunder number
European Research Council310873
Israel Science Foundation
United States-Israel Binational Science Foundation2013284, 2009418


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