Synthesis of soluble myelin-associated glycoprotein in insect and mammalian cells

Paul W. Johnson; John Attia; Christopher D. Richardson; John C. Roder; Robert J. Dunn

doi:10.1016/0378-1119(89)90076-0

Synthesis of soluble myelin-associated glycoprotein in insect and mammalian cells

Paul W. Johnson, John Attia, Christopher D. Richardson, John C. Roder, Robert J. Dunn

Résultat de recherche: Article › examen par les pairs

9 Citations (Scopus)

Résumé

The myelin-associated glycoprotein (MAG) has an extracellular domain containing five sequences which are homologous to the immunoglobulin-fold motif. Adhesive interactions mediated by the MAG extracellular domain are involved in the development of the myelin sheath. The MAG cDNA has been modified to introduce a stop codon immediately before the transmembrane domain. Expression of the modified cDNA in insect cells and murine NIH-3T3 cells resulted in secretion of the soluble MAG extracellular domain. Treatment of soluble MAG with glycopeptidase F and endoglycosidase H showed significant differences in glycosylation for the insect and mammalian cell-expression systems. The soluble form of MAG has been purified from insect-cell supernatants by adsorption to a lentil-lectin support. The soluble MAG will provide a powerful new approach for studies of MAG-adhesive interactions during brain development.

Langue d'origine	English
Pages (de-à)	287-296
Nombre de pages	10
Journal	Gene
Volume	77
Numéro de publication	2
DOI	https://doi.org/10.1016/0378-1119(89)90076-0
Statut de publication	Published - avr. 30 1989
Publié à l'externe	Oui

Note bibliographique

Funding Information:
We thank Dr. Max Summers for providing the insect expression system, Dr. Norman Latov for antibodies and Dr. Tony Pawson for the pECE vector. We thank Dr. Wanda Abramow-Newerly and Martha Garrett for technical assistance and advice, Mike Tropak and Dr. Monique Arquint for plasmids, advice and discussion, and Dr. Reinhart Reithmeier for advice on protein purification. This work was supported by grants from the Medical Research Council of Canada and the Multiple Sclerosis Society of Canada to R.J.D. and J.C.R.

ASJC Scopus Subject Areas

Genetics

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10.1016/0378-1119(89)90076-0

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title = "Synthesis of soluble myelin-associated glycoprotein in insect and mammalian cells",

abstract = "The myelin-associated glycoprotein (MAG) has an extracellular domain containing five sequences which are homologous to the immunoglobulin-fold motif. Adhesive interactions mediated by the MAG extracellular domain are involved in the development of the myelin sheath. The MAG cDNA has been modified to introduce a stop codon immediately before the transmembrane domain. Expression of the modified cDNA in insect cells and murine NIH-3T3 cells resulted in secretion of the soluble MAG extracellular domain. Treatment of soluble MAG with glycopeptidase F and endoglycosidase H showed significant differences in glycosylation for the insect and mammalian cell-expression systems. The soluble form of MAG has been purified from insect-cell supernatants by adsorption to a lentil-lectin support. The soluble MAG will provide a powerful new approach for studies of MAG-adhesive interactions during brain development.",

author = "Johnson, {Paul W.} and John Attia and Richardson, {Christopher D.} and Roder, {John C.} and Dunn, {Robert J.}",

note = "Funding Information: We thank Dr. Max Summers for providing the insect expression system, Dr. Norman Latov for antibodies and Dr. Tony Pawson for the pECE vector. We thank Dr. Wanda Abramow-Newerly and Martha Garrett for technical assistance and advice, Mike Tropak and Dr. Monique Arquint for plasmids, advice and discussion, and Dr. Reinhart Reithmeier for advice on protein purification. This work was supported by grants from the Medical Research Council of Canada and the Multiple Sclerosis Society of Canada to R.J.D. and J.C.R.",

year = "1989",

month = apr,

day = "30",

doi = "10.1016/0378-1119(89)90076-0",

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AU - Johnson, Paul W.

AU - Attia, John

AU - Richardson, Christopher D.

AU - Roder, John C.

AU - Dunn, Robert J.

N1 - Funding Information: We thank Dr. Max Summers for providing the insect expression system, Dr. Norman Latov for antibodies and Dr. Tony Pawson for the pECE vector. We thank Dr. Wanda Abramow-Newerly and Martha Garrett for technical assistance and advice, Mike Tropak and Dr. Monique Arquint for plasmids, advice and discussion, and Dr. Reinhart Reithmeier for advice on protein purification. This work was supported by grants from the Medical Research Council of Canada and the Multiple Sclerosis Society of Canada to R.J.D. and J.C.R.

PY - 1989/4/30

Y1 - 1989/4/30

N2 - The myelin-associated glycoprotein (MAG) has an extracellular domain containing five sequences which are homologous to the immunoglobulin-fold motif. Adhesive interactions mediated by the MAG extracellular domain are involved in the development of the myelin sheath. The MAG cDNA has been modified to introduce a stop codon immediately before the transmembrane domain. Expression of the modified cDNA in insect cells and murine NIH-3T3 cells resulted in secretion of the soluble MAG extracellular domain. Treatment of soluble MAG with glycopeptidase F and endoglycosidase H showed significant differences in glycosylation for the insect and mammalian cell-expression systems. The soluble form of MAG has been purified from insect-cell supernatants by adsorption to a lentil-lectin support. The soluble MAG will provide a powerful new approach for studies of MAG-adhesive interactions during brain development.

AB - The myelin-associated glycoprotein (MAG) has an extracellular domain containing five sequences which are homologous to the immunoglobulin-fold motif. Adhesive interactions mediated by the MAG extracellular domain are involved in the development of the myelin sheath. The MAG cDNA has been modified to introduce a stop codon immediately before the transmembrane domain. Expression of the modified cDNA in insect cells and murine NIH-3T3 cells resulted in secretion of the soluble MAG extracellular domain. Treatment of soluble MAG with glycopeptidase F and endoglycosidase H showed significant differences in glycosylation for the insect and mammalian cell-expression systems. The soluble form of MAG has been purified from insect-cell supernatants by adsorption to a lentil-lectin support. The soluble MAG will provide a powerful new approach for studies of MAG-adhesive interactions during brain development.

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Synthesis of soluble myelin-associated glycoprotein in insect and mammalian cells

Résumé

Note bibliographique

ASJC Scopus Subject Areas

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