Multiple display of catalytic modules on a protein scaffold: nano-fabrication of enzyme particles.
Self assembly is a prerequisite for fabricating nanoscale structures. Here we present a new fusion protein based on the stress-responsive homo-oligomeric protein, SP1. This ring-shaped protein is a highly stable homododecamer, which can be potentially utilized to self-assemble different modules and enzymes in a predicted and oriented manner. For that purpose, a cohesin module (a component of the bacterial cellulosome) was selected, its gene fused in-frame to SP1, and the fusion protein was expressed in Escherichia coli. The cohesin module, specialized to incorporate different enzymes through specific recognition of a dockerin modular counterpart, is used to display new moieties on the SP1 scaffold. The SP1 scaffold displayed 12 active cohesin modules and specific binding to a dockerin-fused cellulase enzyme from Thermobifida fusca. Moreover, we found a significant increase in specific activity of the scaffold-displayed enzymes.
Heyman
Arnon
A
The Robert H Smith Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot, Israel.
Barak
Yoav
Y
Caspi
Jonathan
J
Wilson
David B
DB
Altman
Arie
A
Bayer
Edward A
EA
Shoseyov
Oded
O
D002384:Catalysis; D018797:Cell Cycle Proteins / Q000378:metabolism; D002480:Cellulase / Q000378:metabolism*; D002850:Chromatography, Gel; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D036103:Nanotechnology; D009687:Nuclear Proteins / Q000378:metabolism; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism*; D000097722:Cohesins