The Clostridium cellulovorans cellulosome.
The Clostridium cellulovorans cellulosome is comprised of a large, nonenzymatic scaffolding protein called the cellulose binding protein A (CbpA) and a number of endoglucanases/xylanases. The CbpA contains several functional domains, including a signal peptide, a cellulose binding domain (CBD), a hydrophilic domain (HLD) present four times, and a hydrophobic domain (HBD) present nine times. The functions of the domains were studied by the construction of minigenes containing the putative functional domains and by expression of the minigenes in Escherichia coli. The purified product of the CBD was able to bind to various crystalline forms of cellulose and chitin with a Kd of 1 microM. The binding capacity for CBD was a function of the crystallinity of the cellulose sample. Furthermore, the binding of CBD to Avicel was not inhibited by cellobiose or carboxymethylcellulose, suggesting that the CBD binding target was a three-dimensional structure found only in crystalline forms of cellulose. The HBD was tested for its ability to bind endoglucanases by an interaction Western as well as a sandwich enzyme immunoassay technique. The HBD was able to bind both EngB and EngD, indicating that the HBD contained an endoglucanase binding domain (EBD). Because there are nine EBD domains, it is possible that CbpA can bind up to nine endoglucanases. The role of the HLDs remains elusive. The data indicate that the cellulosome is a complex enzyme containing a scaffolding protein (CbpA) to which is attached a number of endoglucanase molecules. This arrangement allows the complex to bind and degrade crystalline cellulose, which resists degradation by the free forms of cellulosomal endoglucanases.
Doi
R H
RH
Section of Molecular and Cellular Biology, University of California, Davis 95616.
Goldstein
M
M
Hashida
S
S
Park
J S
JS
Takagi
M
M
D000596:Amino Acids / Q000032:analysis; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D002352:Carrier Proteins; D002480:Cellulase / Q000378:metabolism; D002482:Cellulose / Q000378:metabolism*; D003013:Clostridium / Q000235:genetics / Q000378:metabolism*; D005798:Genes, Bacterial; D006026:Glycoside Hydrolases / Q000378:metabolism; D008956:Models, Chemical; D015394:Molecular Structure; D043325:Xylan Endo-1,3-beta-Xylosidase
D000596:Amino Acids; D001426:Bacterial Proteins; D002352:Carrier Proteins; C074074:cellulose binding protein A, Clostridium cellulovorans; D002482:Cellulose; D006026:Glycoside Hydrolases; D043325:Xylan Endo-1,3-beta-Xylosidase; D002480:Cellulase