Heterologous expression of a green fluorescent protein-pertussis toxin S1 subunit fusion construct disrupts calcium channel modulation in rat superior cervical ganglion neurons.
Neurosci Lett
; 271(3): 163-6, 1999 Aug 27.
Article
en En
| MEDLINE
| ID: mdl-10507694
ABSTRACT
The fusion construct pEGFP-PTXS1 was assembled by ligating cDNA encoding the S1 subunit of Bordetella pertussis toxin (PTX) into the plasmid pEGFP-C1 (which codes for enhanced green fluorescent protein). Microinjection of pEGFP-PTXS1 (1-100 ng/microl) into the nucleus of dissociated rat sympathetic ganglion neurons resulted in functional expression as determined from the diffuse green fluorescence and disruption of norepinephrine-mediated N-type Ca2+ channel modulation. The heterologously expressed toxin retained specificity for G alpha(i/o)-dependent pathways as VIP-mediated modulation of N-type Ca2+ channels and muscarine-mediated inhibition of M-type K+ channels persisted in pEGFP-PTXS1 expressing neurons. These data demonstrate that the S1 subunit of PTX is readily expressed in mammalian neurons and remains functional following fusion to the C-terminus of another protein.
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Banco de datos:
MEDLINE
Asunto principal:
Factores de Virulencia de Bordetella
/
Ganglio Cervical Superior
/
Canales de Calcio Tipo N
/
Toxina del Pertussis
/
Proteínas Luminiscentes
/
Neuronas
Límite:
Animals
Idioma:
En
Año:
1999
Tipo del documento:
Article