Purification and crystallization of a multimodular heterotrimeric complex containing both type I and type II cohesin-dockerin interactions from the cellulosome of Clostridium thermocellum.
The multimodular scaffoldin subunit CipA is the central component of the cellulosome, a multienzyme plant cell-wall-degrading complex, from Clostridium thermocellum. It captures secreted cellulases and hemicellulases and anchors the entire complex to the cell surface via high-affinity calcium-dependent interactions between cohesin and dockerin modules termed type I and type II interactions. The crystallization of a heterotrimeric complex comprising the type II cohesin module from the cell-surface protein SdbA, a trimodular C-terminal fragment of the scaffoldin CipA and the type I dockerin module from the CelD cellulase is reported. The crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 119.37, b = 186.31, c = 191.17 A. The crystals diffracted to 2.7 A resolution with four or eight molecules of the ternary protein complex in the asymmetric unit.
Currie
Mark A
MA
Department of Biochemistry, Queen's University, Kingston, ON K7L 3N6, Canada.
Adams
Jarrett J
JJ
Ali
Sabrina
S
Smith
Steven P
SP
Jia
Zongchao
Z
D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D002480:Cellulase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D044603:Cellulosomes / Q000737:chemistry* / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D048013:Acetivibrio thermocellus / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray; D055503:Protein Multimerization; D000097722:Cohesins
D018797:Cell Cycle Proteins; D002480:Cellulase; D002868:Chromosomal Proteins, Non-Histone; D000097722:Cohesins