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Titolo:
Characterization of coated vesicles that participate in endocytic recycling
Autore:
Peters, PJ; Gao, MG; Gaschet, J; Ambach, A; van Donselaar, E; Traverse, JF; Bos, E; Wolffe, EJ; Hsu, VW;
Indirizzi:
Harvard Univ, Brigham & Womens Hosp, Sch Med, Div Rheumatol Allergy & Immunol, Boston, MA 02115 USA Harvard Univ Boston MA USA 02115 Allergy & Immunol, Boston, MA 02115 USA Harvard Univ, Sch Med, Dept Med, Boston, MA 02115 USA Harvard Univ BostonMA USA 02115 Sch Med, Dept Med, Boston, MA 02115 USA Netherlands Canc Inst, Sect Tumor Biol, NL-1066 CX Amsterdam, Netherlands Netherlands Canc Inst Amsterdam Netherlands NL-1066 CX rdam, Netherlands NIAID, Viral Dis Lab, Bethesda, MD 20892 USA NIAID Bethesda MD USA 20892NIAID, Viral Dis Lab, Bethesda, MD 20892 USA
Titolo Testata:
TRAFFIC
fascicolo: 12, volume: 2, anno: 2001,
pagine: 885 - 895
SICI:
1398-9219(200112)2:12<885:COCVTP>2.0.ZU;2-X
Fonte:
ISI
Lingua:
ENG
Soggetto:
ADP-RIBOSYLATION FACTOR; GTP-BINDING PROTEIN; GOLGI MEMBRANES; PLASMA-MEMBRANE; SIGNAL-TRANSDUCTION; NUCLEOTIDE-EXCHANGE; EPITHELIAL-CELLS; PHOSPHOLIPASE-D; ARF FAMILY; RECRUITMENT;
Keywords:
ARF6 coat protein; endocytosis; recycling; vesicles;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
48
Recensione:
Indirizzi per estratti:
Indirizzo: Peters, PJ Harvard Univ, Brigham & Womens Hosp, Sch Med, Div Rheumatol Allergy & Immunol, Boston, MA 02115 USA Harvard Univ Boston MA USA 02115 Immunol, Boston, MA 02115 USA
Citazione:
P.J. Peters et al., "Characterization of coated vesicles that participate in endocytic recycling", TRAFFIC, 2(12), 2001, pp. 885-895

Abstract

While the recycling pathway of endocytosis has been shown to participate in many cellular functions, little is known regarding the transport carriersthat mediate this pathway. In this study, we overexpressed a point mutant of ADP-ribosylation factor 6 (ARF6), that perturbs its GTPase cycle, to accumulate endosome-derived coated vesicles. Characterization by their purification revealed that, upon cell homogenization, these vesicles were mostly aggregated with larger noncoated membranes, and could be released with high-salt treatment. Equilibrium centrifugation revealed that these vesicles hadbuoyant density similar to the COP-coated vesicles. To purify the ARF6-regulated vesicles to homogeneity, enriched fractions from equilibrium centrifugation were subjected to immunoisolation through the hemagglutinin (HA) epitope of the mutant ARF6, by using a newly developed, high-affinity, anti-HA monoclonal antibody. Surface iodination of the purified vesicles revealedmultiple prominent proteins. Immunoblotting with antibodies against subunits of the currently known coat proteins suggested that these vesicles have a novel coat complex. These vesicles are carriers for endocytic recycling, because they are enriched for transferrin receptor and also the v-SNARE cellubrevin that functions in transport from the recycling endosome to the plasma membrane. Thus, we have characterized transport vesicles that participate in endocytic recycling.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/04/20 alle ore 15:06:50