INVESTIGADORES
FOSSATI Carlos Alberto
artículos
Título:
Engineering of a polymeric bacterial protein as a scaffold for the multiple display of peptides
Autor/es:
D A, LAPLAGNE; AS A, ZYLBERMAN ENGINEERING OF A POLYMERIC BACTERI; AINCIART N,; W, STEWARD M; SCIUTTO E,; F G, GOLDBAUM; CARLOS ALBERTO FOSSATI
Revista:
PROTEINS: STRUCTURE, FUNCTION AND GENETICS
Editorial:
WILEY-LISS, DIV JOHN WILEY & SONS INC
Referencias:
Año: 2004 vol. 57 p. 820 - 828
ISSN:
0887-3585
Resumen:
Protein assemblies with a high degree
of repetitiveness and organization are known
to induce strong immune responses. For that reason
they have been postulated for the design of subunit
vaccines by means of protein engineering. The enzyme
lumazine synthase from Brucella spp. (BLS) is
highly immunogenic, presumably owing to its homodecameric
arrangement and remarkable thermodynamic
stability. Structural analysis has shown that
it is possible to insert foreign peptides at the ten
amino terminus of BLS without disrupting its general
folding. These peptides would be displayed to
the immune system in a highly symmetric threedimensional
array. In the present work, BLS has
been used as a protein carrier of foreign peptides.
We have established a modular system to produce
chimeric proteins decorated with ten copies of a
desired peptide as long as 27 residues and have
shown that their folding and stability is similar to
that of the wild-type protein. The knowledge about
the mechanisms of dissociation and unfolding of
BLS allowed the engineering of polyvalent chimeras
displaying different predefined peptides on the same
molecular scaffold. Moreover, the reassembly of
mixtures of chimeras at different steps of the unfolding
process was used to control the stoichiometry
and spatial arrangement for the simultaneous display
of different peptides on BLS. This strategy
would be useful for vaccine development and other
biomedical applications.
Brucella spp. (BLS) is
highly immunogenic, presumably owing to its homodecameric
arrangement and remarkable thermodynamic
stability. Structural analysis has shown that
it is possible to insert foreign peptides at the ten
amino terminus of BLS without disrupting its general
folding. These peptides would be displayed to
the immune system in a highly symmetric threedimensional
array. In the present work, BLS has
been used as a protein carrier of foreign peptides.
We have established a modular system to produce
chimeric proteins decorated with ten copies of a
desired peptide as long as 27 residues and have
shown that their folding and stability is similar to
that of the wild-type protein. The knowledge about
the mechanisms of dissociation and unfolding of
BLS allowed the engineering of polyvalent chimeras
displaying different predefined peptides on the same
molecular scaffold. Moreover, the reassembly of
mixtures of chimeras at different steps of the unfolding
process was used to control the stoichiometry
and spatial arrangement for the simultaneous display
of different peptides on BLS. This strategy
would be useful for vaccine development and other
biomedical applications.