Exploiting the 21st amino acid-purifying and labeling proteins by selenolate targeting.
Nat Methods
; 1(1): 61-6, 2004 Oct.
Article
en En
| MEDLINE
| ID: mdl-15782154
ABSTRACT
Selenium is essential to human life and occurs in selenoproteins as selenocysteine (Sec), the 21st amino acid. The selenium atom endows selenocysteine with unique biochemical properties, including a low pK(a) and a high reactivity with many electrophilic agents. Here we describe the introduction of selenocysteine into recombinant non-selenoproteins produced in Escherichia coli, as part of a small tetrapeptide motif at the C terminus. This selenocysteine-containing motif could subsequently be used as a protein tag for purification of the recombinant protein, selenolate-targeted labeling with fluorescent compounds or radiolabeling with either gamma-emitting (75)Se or short-lived positron emitters such as (11)C. The results presented here thus show how a wide range of biotechnological applications can be developed starting from the insertion of selenocysteine into proteins.
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Base de datos:
MEDLINE
Asunto principal:
Proteínas Recombinantes de Fusión
/
Ingeniería de Proteínas
/
Selenocisteína
/
Proteínas de Escherichia coli
Tipo de estudio:
Evaluation_studies
Idioma:
En
Revista:
Nat Methods
Asunto de la revista:
TECNICAS E PROCEDIMENTOS DE LABORATORIO
Año:
2004
Tipo del documento:
Article