Multiple display of a protein domain on a bacterial polymeric scaffold.
The multiple display of protein domains on polymeric scaffolds is an emerging technology for many applications. BLS is a highly immunogenic protein that has an oligomeric structure formed by a 17.2 kDa subunit arranged as a dimer of pentamers. Here we describe the production as well as the structural, functional, and immunological properties of a 9 kDa double-stranded RNA-binding domain (RBD3) fused to the structure of BLS. We demonstrate that the BLS and RBD3 modules are stably and independently folded in the structure of the chimera and form a decameric structure of 255 kDa as the native BLS oligomers. The polymeric display of RBD3 in the structure of BLS increases the dsRNA binding strength of this domain both in vitro and in vivo, and also enhances its immunogenicity to the point that it breaks the tolerance of mice to the RBD3 self-antigen. Our results underscore the BLS display strategy as a powerful tool for biotechnological and therapeutic applications.
Craig
P O
PO
Fundación Instituto Leloir, Buenos Aires, Argentina.
Berguer
P M
PM
Ainciart
N
N
Zylberman
V
V
Thomas
M G
MG
Martinez Tosar
L J
LJ
Bulloj
A
A
Boccaccio
G L
GL
Goldbaum
F A
FA
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D001665:Binding Sites; D002002:Brucella / Q000201:enzymology; D002942:Circular Dichroism; D003001:Cloning, Molecular; D005260:Female; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D011489:Protein Denaturation; D020836:Protein Structure, Quaternary; D021122:Protein Subunits; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D013816:Thermodynamics
D001426:Bacterial Proteins; D009097:Multienzyme Complexes; D021122:Protein Subunits; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; C044822:6,7-dimethyl-8-ribityllumazine synthase