The murine monoclonal antibody NaM26-4C6 identifies a common structure on band 3 and glycophorin C
| Autor: | M Duk, Marcin Czerwinski, M J Loirat, D Blanchard |
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| Rok vydání: | 1999 |
| Předmět: |
Erythrocytes
medicine.drug_class Immunoblotting Monoclonal antibody Epitope Umbilical Cord Mice Anion Exchange Protein 1 Erythrocyte medicine Glycophorin Animals Humans Glycophorins Peptide sequence Band 3 Mice Inbred BALB C biology Antibodies Monoclonal Hematology Glycophorin C Hemagglutination Tests Molecular biology Immunohistochemistry Protein Structure Tertiary Epitope mapping biology.protein Antibody Epitope Mapping |
| Zdroj: | Transfusion medicine (Oxford, England). 9(1) |
| ISSN: | 0958-7578 |
| Popis: | The murine monoclonal antibody NaM26-4C6 (IgM class), obtained from the splenocytes of a BALB/c mouse immunized with human umbilical cord red blood cells, was characterized by agglutination test and immunoblotting analysis. The structure of the NaM26-4C6 epitope was further elucidated by using a series of peptides synthesized on pins. The antibody agglutinated untreated and chymotrypsin-treated but not trypsin- or neuraminidase-treated human erythrocytes. Agglutination-inhibition test demonstrated that the antibody recognizes an epitope located on the N-terminal trypsin-sensitive portion of glycophorin C. The antibody bound on immunoblots to glycophorin C, and also to the band 3 protein and its 69-kDa N-terminal fragment but did not bind to desialylated and de-O-glycosylated glycophorin C. Peptide mapping allowed localization of the binding site on the 23-kDa N-terminal intracellular peptide of band 3. The antibody binds to the amino-acid sequences 22EDPDIP27 of band 3 protein and 15SLEPDPGM22 of glycophorin C, and residues D and P were found to be essential. The new epitope identified by NaM26-4C6 corresponds to a linear amino acid sequence located on the N-terminal intracellular portion of band 3 and to a more complex structure involving oligosaccharide chains on the N-terminal extracellular domain of GPC. |
| Databáze: | OpenAIRE |
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