Localization of specific epitopes on human microtubule-associated protein 2
| Autor: | L. I. Binder, Nellie Kalcheva, Bridget Shafit-Zagardo, J. S. Albala |
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| Rok vydání: | 1994 |
| Předmět: |
Microtubule-associated protein
Recombinant Fusion Proteins Blotting Western Gene Expression Biology Biochemistry Epitope Cellular and Molecular Neuroscience Epitopes Neuroblastoma Western blot Microtubule-associated protein 2 medicine Escherichia coli Tumor Cells Cultured Humans Phosphorylation chemistry.chemical_classification medicine.diagnostic_test Antibodies Monoclonal Alkaline Phosphatase Molecular biology Fusion protein Amino acid Molecular Weight Epitope mapping Tubulin chemistry biology.protein Electrophoresis Polyacrylamide Gel Microtubule-Associated Proteins |
| Zdroj: | Journal of neurochemistry. 63(6) |
| ISSN: | 0022-3042 |
| Popis: | Microtubule-associated protein 2 (MAP-2) is an abundant neuronal cytoskeletal protein that binds to tubulin and stabilizes microtubules. Using fusion protein constructs we have defined the epitopes of 10 monoclonal antibodies (mAbs) to discrete regions of human MAP-2. Proteins were expressed in pATH vectors. After electrophoresis, immunoblotting was performed. By western blot analysis five of the mAbs (AP-14, AP-20, AP-21, AP-23, and AP-25) share epitopes with only the high molecular weight isoforms (MAP-2a, MAP-2b); two of the mAbs (AP-18 and tau 46) recognize MAP-2a, MAP-2b, and MAP-2c. Although AP-18 immunoreactivity was detected within heat-stable protein homogenates isolated from a human neuroblastoma cell line MSN, fusion protein constructs encompassing human MAP-2 were negative, suggesting that the AP-18 epitope is phosphorylated. Furthermore, AP-18 immunoreactivity was lost after alkaline phosphatase treatment of heat-stable protein preparations from MSN cells. Four of the mAbs (322, 636, 635, and 39) recognize epitopes located within amino acids 169–219 of human MAP-2. AP-21 maps to a region between amino acids 553 and 645. AP-23 maps between amino acids 645 and 993, whereas AP-20, AP-14, and AP-25 map between amino acids 995 and 1332. Expression of the region of MAP-2 between amino acids 1787 and 1824 was positive to tau 46. |
| Databáze: | OpenAIRE |
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