Probing Ca2+-binding capability of viral proteins with the EF-hand motif by grafting approach.
Methods Mol Biol
; 963: 37-53, 2013.
Article
em En
| MEDLINE
| ID: mdl-23296603
ABSTRACT
Ca(2+) is implicated in almost every step of the life cycle of viruses, including virus entry into host cells, virus replication, virion assembly, maturation, and release. However, due to the lack of prediction algorithms and rigorous validation methods, only limited cases of viral Ca(2+)-binding sites are reported. Here, we introduce a method to predict continuous EF-hand or EF-hand-like motifs in the viral genomes based on their primary sequences. We then introduce a grafting approach, and the use of luminescence resonance energy transfer and Ca(2+) competition assay for experimental verification of predicted Ca(2+)-binding sites. This protocol will be valuable for the prediction and identification of unknown Ca(2+)-binding sites in virus.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteínas Virais
/
Cálcio
/
Biologia Computacional
/
Motivos EF Hand
Tipo de estudo:
Prognostic_studies
Idioma:
En
Revista:
Methods Mol Biol
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2013
Tipo de documento:
Article
País de afiliação:
Estados Unidos