Cohesin-dockerin microarray: Diverse specificities between two complementary families of interacting protein modules.
The cellulosome is an intricate multienzyme complex, designed for efficient degradation of plant cell wall polysaccharides, notably cellulose. The supramolecular cellulosome architecture in different bacteria is the consequence of the types and specificities of the interacting cohesin and dockerin modules, borne by the different cellulosomal subunits. In this study, we describe a microarray system for determining cohesin-dockerin specificity, which allows global comparison among the interactions between various members of these two complementary families of interacting protein modules. Matching recombinant fusion proteins were prepared that contained one of the interacting modules: cohesins were joined to an appropriate cellulose-binding module (CBM) and the dockerins were fused to a thermostable xylanase that served to enhance expression and proper folding. The CBM-fused cohesins were immobilized on cellulose-coated glass slides, to which xylanase-fused dockerin samples were applied. Knowledge of the specificity characteristics of native and mutated members of the cohesin and dockerin families provides insight into the architecture of the parent cellulosome and allows selection of suitable cohesin-dockein pairs for biotechnological and nanotechnological application. Using this approach, extensive cross-species interaction among type-II cohesins and dockerins is shown for the first time. Selective intraspecies binding of an archaeal dockerin to two complementary cohesins is also demonstrated.
Haimovitz
Rachel
R
Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot, Israel.
Barak
Yoav
Y
Morag
Ely
E
Voronov-Goldman
Milana
M
Shoham
Yuval
Y
Lamed
Raphael
R
Bayer
Edward A
EA
D001105:Archaea / Q000737:chemistry; D019843:Archaeal Proteins / Q000378:metabolism*; D001419:Bacteria / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism*; D044603:Cellulosomes / Q000737:chemistry; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D009097:Multienzyme Complexes; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism*; D040081:Protein Array Analysis; D000097722:Cohesins
D019843:Archaeal Proteins; D001426:Bacterial Proteins; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D008565:Membrane Proteins; D009097:Multienzyme Complexes; D009687:Nuclear Proteins; D000097722:Cohesins