Journal article
Specificity of human IgM M-proteins that bind to myelin-associated glycoprotein: peptide mapping, deglycosylation, and competitive binding studies
The Journal of immunology (1950), Vol.133(5), pp.2509-2512
1984
DOI: 10.4049/jimmunol.133.5.2509
PMID: 6207233
Abstract
Ten patients with neuropathy and IgM M-proteins that bind to the myelin-associated glycoprotein (MAG) were studied to determine whether the M-proteins bind to common regions of MAG and whether the reactive determinants contain carbohydrate residues. The M-protein of one patient was biotinylated, and binding to human MAG was quantitated by enzyme-linked immunosorbent assay (ELISA) by using avidin-biotin-peroxidase complexes. Serum from the same patient and nine others, but not from controls, competed with the biotinylated M-protein for binding to human MAG. Bovine MAG was digested with staph protease or cleaved with cyanogen bromide, and the resultant fragments were separated by electrophoresis and were transferred onto nitrocellulose sheets. Serum from all patients immunostained the peptide fragments identically. Bovine MAG was deglycosylated by trifluoromethanesulfonic acid, and binding of the M-proteins to MAG and to deglycosylated MAG was tested by immunoblotting. None of the patient's M-proteins bound to deglycosylated MAG. Deglycosylated MAG was visualized by using a mouse monoclonal antibody, GEN-S3, directed at the polypeptide core of MAG. The effectiveness of deglycosylation was ascertained by electrophoresis and by binding of biotinylated concanavalin A. These data and the observed identical species specificity of the M-proteins suggest that the respective anti-MAG M-proteins all bind to the same region in MAG and that the reactive determinants may contain carbohydrate moieties.
Details
- Title: Subtitle
- Specificity of human IgM M-proteins that bind to myelin-associated glycoprotein: peptide mapping, deglycosylation, and competitive binding studies
- Creators
- Michael E Shy - Columbia univ., coll. physicians surgeons, New York NY 10032, United StatesThomas Vietorisz - Columbia univ., coll. physicians surgeons, New York NY 10032, United StatesEduardo Nobile-Orazio - Columbia univ., coll. physicians surgeons, New York NY 10032, United StatesNorman Latov - Columbia univ., coll. physicians surgeons, New York NY 10032, United States
- Resource Type
- Journal article
- Publication Details
- The Journal of immunology (1950), Vol.133(5), pp.2509-2512
- Publisher
- American Association of Immunologists; Bethesda, MD
- DOI
- 10.4049/jimmunol.133.5.2509
- PMID
- 6207233
- ISSN
- 0022-1767
- eISSN
- 1550-6606
- Language
- English
- Date published
- 1984
- Academic Unit
- Neurology; Molecular Physiology and Biophysics; Stead Family Department of Pediatrics; Iowa Neuroscience Institute
- Record Identifier
- 9984020740602771
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