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Titolo:
RECONSTITUTION OF 5'-NUCLEOTIDASE OF BULL SEMINAL PLASMA IN SPIN-LABELED LIPOSOMES
Autore:
FINI C; THUONG VD; ALIANTE M; FLORIDI A; CANNISTRARO S;
Indirizzi:
UNIV PERUGIA,DIPARTIMENTO BIOL CELLULARE & MOLEC,VIA DEL GIOCHETTO 6 I-06126 PERUGIA ITALY
Titolo Testata:
The Journal of membrane biology
fascicolo: 1, volume: 142, anno: 1994,
pagine: 137 - 144
SICI:
0022-2631(1994)142:1<137:RO5OBS>2.0.ZU;2-J
Fonte:
ISI
Lingua:
ENG
Soggetto:
SOLUBLE 5'-NUCLEOTIDASE; LIVER 5'-NUCLEOTIDASE; ANCHORED PROTEINS; CHICKEN GIZZARD; RAT-HEART; PURIFICATION; MEMBRANE; IDENTIFICATION; EXISTENCE; RESONANCE;
Keywords:
5'-NUCLEOTIDASE; LIPOSOMES; GPI-ANCHORED PROTEINS; ECTOENZYMES; ELECTRON SPIN PROBES; DISULFIDE BRIDGES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
33
Recensione:
Indirizzi per estratti:
Citazione:
C. Fini et al., "RECONSTITUTION OF 5'-NUCLEOTIDASE OF BULL SEMINAL PLASMA IN SPIN-LABELED LIPOSOMES", The Journal of membrane biology, 142(1), 1994, pp. 137-144

Abstract

Seminal plasma separated from freshly ejaculated bull semen contains vesicles with a 5'-nucleotidase activity incorporated as an ectoenzymeanchored by glycosyl phosphatidylinositol (GPI). After its extractionfrom bull seminal plasma vesicles, the protein was purified and reconstituted into hen egg yolk lecithin liposomes obtained through prolonged dialysis of buffered n-octylglucoside detergent solutions of lipid,protein and various effecters against detergent-free solutions. Gel filtration experiments showed that the enzyme incorporated into liposomes in a dimeric form with its two subunits linked by disulfide bridges. In the presence of reduced glutathione, the protein dissociated intomonomers and failed to incorporate into liposomes. Electron spin resonance (ESR) experiments, performed with liposomes containing electron spin labels localized at the hydrophilic lipid headgroups (5-doxyl stearic acid) or in the hydrophobic lipid hydrocarbon chains (16-doxyl stearic acid), demonstrated that the incorporation of 5'-nucleotidase resulted in the immobilization of the spin probes. Furthermore, the spectral parameters obtained before and after treatment of 5'-nucleotidase-containing liposomes with phosphatidylinositol-specific phospholipaseC (PI-PLC) indicated that the liposome membrane bilayer did not contain protein segments. This supports the well-known ecto-localization of5'-nucleotidase and rules out a previously reported possibility of a proteic transmembrane anchoring of the enzyme.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 02/12/20 alle ore 14:41:42