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Determinants of the intracellular fate of truncated forms of the platelet glycoproteins IIb and IIIa.
- Source :
-
The Journal of biological chemistry [J Biol Chem] 1993 Feb 15; Vol. 268 (5), pp. 3580-5. - Publication Year :
- 1993
-
Abstract
- The platelet glycoproteins GPIIb and GPIIIa are integral membrane proteins and form calcium-dependent heterodimers in the endoplasmic reticulum (ER). In the absence of heterodimer formation, GPIIb and GPIIIa are retained in the ER and degraded. To produce soluble forms of these proteins, we truncated each at a site just proximal to its transmembrane anchor and expressed the mutants in COS-1 cells. We found that both truncated GPIIIa (GPIIIatr) and GPIIIatr were secreted by the transfected cells. However, GPIIbtr was retained by the cells and was immunoprecipitated as a doublet with a 115,000 molecular weight protein. Incubation of transfected cells with the calcium ionophore A23187 or the calcium chelator 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl) ester (BAPTA-AM) failed to induce appreciable GPIIbtr secretion, suggesting that formation of intracellular calcium complexes was not a factor in GPIIbtr retention. Further, immunoblotting of immunoprecipitated GPIIbtr and GPIIIatr revealed that the chaperone binding protein (BiP) was associated with each, arguing that BiP alone was not responsible for GPIIbtr retention. These studies indicate that the intracellular retention of GPIIIa involves sequences located in the transmembrane or cytoplasmic domains of the molecule. GPIIb contains an additional retention signal located in the extracellular portion of the molecule whose effect is abrogated by formation of a GPIIb-IIIa heterodimer. This signal may be involved in the fate of nascent GPIIb monomers and the generation of correctly configured GPIIb-IIIa heterodimers.
- Subjects :
- Amino Acid Sequence
Animals
Base Sequence
Calcimycin pharmacology
Cell Line
Cell Membrane metabolism
DNA genetics
Egtazic Acid analogs & derivatives
Egtazic Acid pharmacology
Endoplasmic Reticulum metabolism
Kinetics
Macromolecular Substances
Molecular Sequence Data
Plasmids
Platelet Membrane Glycoproteins genetics
Platelet Membrane Glycoproteins isolation & purification
Recombinant Proteins isolation & purification
Recombinant Proteins metabolism
Transfection
Mutagenesis, Site-Directed
Platelet Membrane Glycoproteins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0021-9258
- Volume :
- 268
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- The Journal of biological chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 8429034