High-throughput screening of cohesin mutant libraries on cellulose microarrays

Michal Slutzki*, Vered Ruimy, Ely Morag, Yoav Barak, Rachel Haimovitz, Raphael Lamed, Edward A. Bayer

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review


The specificity of cohesin-dockerin interactions is critically important for the assembly of cellulosomal enzymes into the multienzyme cellulolytic complex (cellulosome). In order to investigate the origins of the observed specificity, a variety of selected amino acid positions at the cohesin-dockerin interface can be subjected to mutagenesis, and a library of mutants can be constructed. In this chapter, we describe a protein-protein microarray technique based on the high affinity of a carbohydrate-binding module (CBM), attached to mutant cohesins. Using cellulose-coated glass slides, libraries of mutants can be screened for binding to complementary partners. The advantages of this tool are that crude cell lysate can be used without additional purification, and the microarray can be used for screening both large libraries as initial scanning for "positive" plates, and for small libraries, wherein individual colonies are printed on the slide. Since the time-consuming step of purifying proteins can be circumvented, the approach is also appropriate for providing molecular insight into the multicomponent organization of complex cellulosomes.

Original languageEnglish
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Number of pages11
StatePublished - 2012

Publication series

NameMethods in Enzymology
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988


  • CBM
  • Cellulosome
  • Cohesin-dockerin interaction
  • Protein microarray


Dive into the research topics of 'High-throughput screening of cohesin mutant libraries on cellulose microarrays'. Together they form a unique fingerprint.

Cite this