Preparation of 2-azidoadenosine 3',5'-[5'-32P]bisphosphate for incorporation into transfer RNA. Photoaffinity labeling of Escherichia coli ribosomes.
FEBS Lett
; 245(1-2): 9-13, 1989 Mar 13.
Article
en En
| MEDLINE
| ID: mdl-2647526
2-Azidoadenosine was synthesized from 2-chloroadenosine by sequential reaction with hydrazine and nitrous acid and then bisphosphorylated with pyrophosphoryl chloride to form 2-azidoadenosine 3',5'-bisphosphate. The bisphosphate was labeled in the 5'-position using the exchange reaction catalyzed by T4 polynucleotide kinase in the presence of [gamma-32P]ATP. Polynucleotide kinase from a T4 mutant which lacks 3'-phosphatase activity (ATP:5'-dephosphopolynucleotide 5'-phosphotransferase, EC 2.7.1.78) was required to facilitate this reaction. 2-Azidoadenosine 3',5'-[5'-32P]bisphosphate can serve as an efficient donor in the T4 RNA ligase reaction and can replace the 3'-terminal adenosine of yeast tRNAPhe with little effect on the amino acid acceptor activity of the tRNA. In addition, we show that the modified tRNAPhe derivative can be photochemically cross-linked to the Escherichia coli ribosome.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Ribosomas
/
Azidas
/
ARN de Transferencia
/
Adenosina Difosfato
/
Escherichia coli
Idioma:
En
Revista:
FEBS Lett
Año:
1989
Tipo del documento:
Article
Pais de publicación:
Reino Unido