In vivo incorporation of non-canonical amino acids by using the chemical aminoacylation strategy: a broadly applicable mechanistic tool.
Chembiochem
; 15(12): 1710-20, 2014 Aug 18.
Article
em En
| MEDLINE
| ID: mdl-24990307
ABSTRACT
We describe a strategy for incorporating non-canonical amino acids site-specifically into proteins expressed in living cells, involving organic synthesis to chemically aminoacylate a suppressor tRNA, protein expression in Xenopus oocytes, and monitoring protein function, primarily by electrophysiology. With this protocol, a very wide range of non-canonical amino acids can be employed, allowing both systematic structure-function studies and the incorporation of reactive functionalities. Here, we present an overview of the methodology and examples meant to illustrate the versatility and power of the method as a tool for investigating protein structure and function.
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Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Aminoacil-RNA de Transferência
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Proteínas
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Aminoacilação de RNA de Transferência
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Aminoácidos
Limite:
Animals
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Humans
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article