Investigating the refolding pathway of human acidic fibroblast growth factor (hFGF-1) from the residual structure(s) obtained by denatured-state hydrogen/deuterium exchange.
Biophys J
; 100(1): 154-64, 2011 Jan 05.
Article
em En
| MEDLINE
| ID: mdl-21190667
ABSTRACT
Human fibroblast growth factor 1 (hFGF-1) consists of 12 anti-parallel ß-strands arranged into a ß-trefoil architecture. We directly measured hydrogen/deuterium exchange rates on the urea-denatured hFGF-1 to obtain the information with regard to the persistent residual interaction(s) in the unfolded hFGF-1. Thirty-eight residues whose heteronuclear single quantum coherence cross-peaks can be observed after exchange show higher protections than those predicted for the same residues in a random coil conformation, suggesting the existence of residual structure(s). The urea-denaturation of hFGF-1 tested by both circular dichroism and fluorescence spectroscopy indicated that the unfolding process is a cooperative two-state process and that the residual structures observed did not originate from the existence of a partially structured intermediate. The coincident disappearance of the native heteronuclear single quantum coherence cross-peaks during the urea-denaturation process suggests that the residual structures observed contain no nativelike interactions. The protected residues (fold ons) in the urea-denatured state are mostly those that exchange slowly in the native state H/D exchange. The distribution of these fold ons in the native structure of hFGF-1 suggests that the refolding starts by collisions between the residual structures (microdomains) between the ß-strands VI and VII, and between the ß-strands II and III, which appear to be two independent refolding coordinates during the refolding process.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Desnaturação Proteica
/
Fator 1 de Crescimento de Fibroblastos
/
Medição da Troca de Deutério
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Redobramento de Proteína
/
Aminoácidos
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article