Site-Specific Labelling of Native Mammalian Proteins for Single-Molecule FRET Measurements.
Chembiochem
; 19(8): 780-783, 2018 04 16.
Article
em En
| MEDLINE
| ID: mdl-29394002
ABSTRACT
Human cells are complex entities in which molecular recognition and selection are critical for cellular processes often driven by structural changes and dynamic interactions. Biomolecules appear in different chemical states, and modifications, such as phosphorylation, affect their function. Hence, using proteins in their chemically native state in biochemical and biophysical assays is essential. Single-molecule FRET measurements allow exploration of the structure, function and dynamics of biomolecules but cannot be fully exploited for the human proteome, as a method for the site-specific coupling of organic dyes into native, non-recombinant mammalian proteins is lacking. We address this issue showing the site-specific engineering of fluorescent dyes into human proteins on the basis of bioorthogonal reactions. We show the applicability of the method to study functional and post-translationally modified proteins on the single-molecule level, among them the hitherto inaccessible human Argonauteâ
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas
/
Transferência Ressonante de Energia de Fluorescência
Limite:
Humans
Idioma:
En
Ano de publicação:
2018
Tipo de documento:
Article