Localization of specific epitopes on human microtubule-associated protein 2

N. Kalcheva, J. S. Albala, L. I. Binder, Bridget Shafit-Zagardo

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)2336-2341
Number of pages6
JournalJournal of Neurochemistry
Volume63
Issue number6
StatePublished - Dec 1994

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Epitopes
Amino Acids
Monoclonal Antibodies
Microtubule-Associated Proteins
Proteins
Fusion reactions
Hot Temperature
Cytoskeletal Proteins
Tubulin
Electrophoresis
Neuroblastoma
Immunoblotting
Microtubules
Alkaline Phosphatase
Protein Isoforms
Molecular Weight
Western Blotting
Molecular weight
Cells
human MAP2 protein

Keywords

  • Epitope mapping
  • Expression vectors
  • Human brain
  • Microtubule-associated protein
  • Microtubule-associated protein 2

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Localization of specific epitopes on human microtubule-associated protein 2. / Kalcheva, N.; Albala, J. S.; Binder, L. I.; Shafit-Zagardo, Bridget.

In: Journal of Neurochemistry, Vol. 63, No. 6, 12.1994, p. 2336-2341.

Research output: Contribution to journalArticle

Kalcheva, N. ; Albala, J. S. ; Binder, L. I. ; Shafit-Zagardo, Bridget. / Localization of specific epitopes on human microtubule-associated protein 2. In: Journal of Neurochemistry. 1994 ; Vol. 63, No. 6. pp. 2336-2341.
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