title: Multiple display of catalytic modules on a protein scaffold: nano-fabrication of enzyme particles.
abstract: 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.
authors.affiliation: 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.
authors.forename: Arnon
authors.initials: A
authors.lastname: Heyman
authors.forename: Yoav
authors.initials: Y
authors.lastname: Barak
authors.forename: Jonathan
authors.initials: J
authors.lastname: Caspi
authors.forename: David B
authors.initials: DB
authors.lastname: Wilson
authors.forename: Arie
authors.initials: A
authors.lastname: Altman
authors.forename: Edward A
authors.initials: EA
authors.lastname: Bayer
authors.forename: Oded
authors.initials: O
authors.lastname: Shoseyov
mesh_terms: 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