Einzeltreffer — DigiBib

The yeast Saccharomyces cerevisiae was examined as host for heterologous expression of the G protein-coupled VPAC1 receptor. Rat VPAC1 receptor cDNA and two chimeric constructs encoding the yeast mating factor pre-pro alpha-leader peptide fused in-frame to rat VPAC1 receptor were expressed in yeast cells under control of a galactose inducible promoter. The rat VPAC1 receptor was fused to the HSV tag epitope to ensure proper immunological detection of the receptor. Crucial conditions for high-level expression of active rat VPAC1 receptor included growth in amino acid supplemented minimal medium, fusion to the yeast alpha-leader peptide and a temperature shift from 30 degrees C to 15 degrees C before promoter induction. Western blotting showed that the expressed receptor was highly glycosylated and a band of 47 kDa was obtained upon endoglycosidase H treatment. Binding with radioiodinated vasoactive intestinal polypeptide revealed a KD of 2.5 nM and an IC50 of 15 nM when displacing with unlabeled vasoactive intestinal polypeptide. VPAC1 receptor density quantified by Western blotting was 510 pmol/mg membrane protein of which only 66 pmol/mg were able to bind vasoactive intestinal polypeptide.

Titel:	Functional expression of rat VPAC1 receptor in Saccharomyces cerevisiae.
Autor/in / Beteiligte Person:	Hansen, MK ; Tams, JW ; Fahrenkrug, J ; Pedersen, PA
Zeitschrift:	Receptors & channels, Jg. 6 (1999), Heft 4, S. 271
Veröffentlichung:	London : Taylor & Francis ; <i>Original Publication</i>: Yverdon, Switzerland : Harwood Academic Publishers : Distributed by STBS, c1993-, 1999
Medientyp:	academicJournal
ISSN:	1060-6823 (print)
Schlagwort:	Animals Binding, Competitive Cloning, Molecular Culture Media pharmacology DNA, Complementary genetics Fungal Proteins genetics GTP-Binding Proteins metabolism Gene Expression Regulation, Fungal drug effects Glycosylation Promoter Regions, Genetic Protein Precursors genetics Protein Processing, Post-Translational Rats Receptors, Vasoactive Intestinal Peptide genetics Receptors, Vasoactive Intestinal Polypeptide, Type I Recombinant Fusion Proteins biosynthesis Vasoactive Intestinal Peptide metabolism Receptors, Vasoactive Intestinal Peptide biosynthesis Saccharomyces cerevisiae metabolism Saccharomyces cerevisiae Proteins
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article; Research Support, Non-U.S. Gov't Language: English [Recept Channels] 1999; Vol. 6 (4), pp. 271-81. MeSH Terms: Saccharomyces cerevisiae Proteins* ; Receptors, Vasoactive Intestinal Peptide / biosynthesis ; Saccharomyces cerevisiae / metabolism ; Animals ; Binding, Competitive ; Cloning, Molecular ; Culture Media / pharmacology ; DNA, Complementary / genetics ; Fungal Proteins / genetics ; GTP-Binding Proteins / metabolism ; Gene Expression Regulation, Fungal / drug effects ; Glycosylation ; Promoter Regions, Genetic ; Protein Precursors / genetics ; Protein Processing, Post-Translational ; Rats ; Receptors, Vasoactive Intestinal Peptide / genetics ; Receptors, Vasoactive Intestinal Polypeptide, Type I ; Recombinant Fusion Proteins / biosynthesis ; Vasoactive Intestinal Peptide / metabolism Substance Nomenclature: 0 (Culture Media) ; 0 (DNA, Complementary) ; 0 (Fungal Proteins) ; 0 (MF(ALPHA)1 protein, S cerevisiae) ; 0 (Protein Precursors) ; 0 (Receptors, Vasoactive Intestinal Peptide) ; 0 (Receptors, Vasoactive Intestinal Polypeptide, Type I) ; 0 (Recombinant Fusion Proteins) ; 0 (Saccharomyces cerevisiae Proteins) ; 37221-79-7 (Vasoactive Intestinal Peptide) ; EC 3.6.1.- (GTP-Binding Proteins) Entry Date(s): Date Created: 19990721 Date Completed: 19990923 Latest Revision: 20200825 Update Code: 20231215

Klicken Sie ein Format an und speichern Sie dann die Daten oder geben Sie eine Empfänger-Adresse ein und lassen Sie sich per Email zusenden.

BibTeX Citavi, JabRef, u.a.
(Literaturverwaltung)

PDF kein Volltext!
(Merkzettel, Notizen)

RIS Endnote, Citavi u.a.
(Literaturverwaltung)

MODS
(XML zur Weiterverarbeitung)

oder

Wählen Sie das für Sie passende Zitationsformat und kopieren Sie es dann in die Zwischenablage, lassen es sich per Mail zusenden oder speichern es als PDF-Datei.

Gewünschter Zitations-Stil:

oder

Bitte prüfen Sie, ob die Zitation formal korrekt ist, bevor Sie sie in einer Arbeit verwenden. Benutzen Sie gegebenenfalls den "Exportieren"-Dialog, wenn Sie ein Literaturverwaltungsprogramm verwenden und die Zitat-Angaben selbst formatieren wollen.

Functional expression of rat VPAC1 receptor in Saccharomyces cerevisiae.