INVESTIGADORES
FERREIRO Diego Ulises
artículos
Título:
A protein-DNA binding mechanism proceeds through multi-state or two-state parallel pathways.
Autor/es:
FERREIRO DU, DE PRAT-GAY G.
Revista:
JOURNAL OF MOLECULAR BIOLOGY
Referencias:
Año: 2003 vol. 331 p. 89 - 99
ISSN:
0022-2836
Resumen:
The DNA-binding mechanism of the dimeric C-terminal domain of the
papillomavirus E2 protein with its specific DNA target was investigated
and shown to proceed through two parallel pathways. A sequential
multi-step reaction is initiated by the diffusion-controlled formation
of an encounter complex, with no evidence of base sequence
discrimination capacity. Following a substantial conformational
rearrangement of the protein, a solvent exclusion step leading to the
formation of a final protein-DNA complex was identified. This last step
involves the largest burial of surface area from the interface and
involves the consolidation of the direct readout of the DNA bases.
Double-jump stopped-flow experiments allowed us to characterize the
sequence of events and demonstrated that a fast-formed consolidated
complex can take place through a parallel route. We present the
simplest model for the overall mechanism with a description of all the
intermediate species in energetic terms.