Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
Enhanced fluorescence resonance energy transfer between spectral variants of green fluorescent protein through zinc-site engineering
Autore:
Jensen, KK; Martini, L; Schwartz, TW;
Indirizzi:
Univ Copenhagen, Panum Inst, Dept Pharmacol, Mol Pharmacol Lab, DK-2200 Copenhagen, Denmark Univ Copenhagen Copenhagen Denmark DK-2200 , DK-2200 Copenhagen, Denmark
Titolo Testata:
BIOCHEMISTRY
fascicolo: 4, volume: 40, anno: 2001,
pagine: 938 - 945
SICI:
0006-2960(20010130)40:4<938:EFRETB>2.0.ZU;2-U
Fonte:
ISI
Lingua:
ENG
Soggetto:
TACHYKININ NK-1 RECEPTOR;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
20
Recensione:
Indirizzi per estratti:
Indirizzo: Schwartz, TW Univ Copenhagen, Panum Inst, Dept Pharmacol, Mol Pharmacol Lab, Blegdamsvej 3C, DK-2200 Copenhagen, Denmark Univ Copenhagen Blegdamsvej 3C Copenhagen Denmark DK-2200 rk
Citazione:
K.K. Jensen et al., "Enhanced fluorescence resonance energy transfer between spectral variants of green fluorescent protein through zinc-site engineering", BIOCHEM, 40(4), 2001, pp. 938-945

Abstract

Although spectral variants of GFP should in theory be suited for fluorescence resonance energy transfer (FRET) and therefore suited for studies of protein-protein interactions, the unfavorable location of the fluorophore 15 Angstrom deep inside the GFP molecule has especially impaired this application. Here, metal-ion site engineering around the dimerization interface known from the X-ray structure of OFF is applied to the cyan and the yellow spectral variant of GFP to stabilize the heterodimeric form of these molecules and thereby increase FRET signaling. The FRET signal, determined as the ratio between the maximal emission for the yellow variant, 530 nm, and the cyan variant, 475 nm, during excitation of the cyan variant at 433 nm was increased up to 8-10-fold in the presence of 10(-4) M ZnCl2 by engineering oftwo symmetric metal-ion sites being either bidentate or tridentate. A similar increase in FRET signaling was however obtained in a pair of molecules in which a single bidentate metal-ion site was generated by introducing a zinc-binding residue in each of the two spectral variants of GFP and therefore creating an obligate heterodimeric pair. It is concluded that FRET signaling between spectral variants of GFP can be increased by stabilizing dimerformation and especially by favoring heterodimer formation in this case performed by metal-ion site engineering.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 28/11/20 alle ore 03:11:16