Segmental isotopic labeling by asparaginyl endopeptidase-mediated protein ligation.
J Biomol NMR
; 71(4): 225-235, 2018 Aug.
Article
em En
| MEDLINE
| ID: mdl-29536230
ABSTRACT
Segmental isotopic labeling can facilitate NMR studies of large proteins, multi-domain proteins, and proteins with repetitive sequences by alleviating NMR signal overlaps. Segmental isotopic labeling also allows us to investigate an individual domain in the context of a full-length protein by NMR. Several established methods are available for segmental isotopic labeling such as intein-mediated ligation, but each has specific requirements and limitations. Here, we report an enzymatic approach using bacterially produced asparagine endopeptidase from Oldenlandia affinis for segmental isotopic labeling of a protein with repetitive sequences, a designed armadillo repeat protein, by overcoming some of the shortcomings of enzymatic ligation for segmental isotopic labeling.
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Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Cisteína Endopeptidases
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Ressonância Magnética Nuclear Biomolecular
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Marcação por Isótopo
Idioma:
En
Revista:
J Biomol NMR
Assunto da revista:
BIOLOGIA MOLECULAR
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DIAGNOSTICO POR IMAGEM
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MEDICINA NUCLEAR
Ano de publicação:
2018
Tipo de documento:
Article
País de afiliação:
Finlândia