Reverse-phase high-performance liquid chromatography of nerve growth factor receptor-like proteins identified with monoclonal antibodies

D. E. Shan, C. E. Beck, K. Werrbach-Perez, J. R. Perez-Polo

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Human neuroblastoma SK-N-SH-SY5Y (SY5Y) and rat pheochromocytoma PC12 cells are model cell lines used in the study of nerve growth factor (NGF) effect. The effects of NGF are initiated by binding to cell surface receptors (NGFR). The amino acid sequence for NGFR has been deduced based on the identification of a single gene for NGFR. However, there are two kinds of NGF binding activities and several reported molecular weights of NGFR. We report here on the demonstration of NGFR-like proteins from PC12 and SY5Y cells by sequential lectin chromatography, reverse-phase HPLC, and SDS-PAGE analysis of immunoprecipitates obtained with NGFR-specific monoclonal antibodies. For both human and rodent NGFR, there was a tendency for the higher molecular-weight species of NGFR-like proteins to be eluted in more hydrophobic fractions. Also, the expression of different species of NGFR could be modified by treatment with retinoic acid (RA). These results are consistent with the hypothesis that the different molecular species of NGFR may result from the generation of a truncated form of NGFR, the presence of sugar residues on the NGFR protein, dimer formation between NGFR, or the association of NGFR with a receptor-associated protein.

Original languageEnglish (US)
Pages (from-to)620-632
Number of pages13
JournalJournal of Neuroscience Research
Volume27
Issue number4
DOIs
StatePublished - 1990
Externally publishedYes

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Nerve Growth Factor Receptor
Nerve Growth Factor
Reverse-Phase Chromatography
PC12 Cells
High Pressure Liquid Chromatography
Monoclonal Antibodies
Molecular Weight
Proteins
Cell Surface Receptors
Pheochromocytoma
Tretinoin
Neuroblastoma
Lectins
Polyacrylamide Gel Electrophoresis
Amino Acid Sequence
Rodentia
Cell Line
Genes

Keywords

  • hydrophobicity
  • NGF
  • NGF receptor
  • purification
  • receptor-associated protein
  • reverse-phase HPLC

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Reverse-phase high-performance liquid chromatography of nerve growth factor receptor-like proteins identified with monoclonal antibodies. / Shan, D. E.; Beck, C. E.; Werrbach-Perez, K.; Perez-Polo, J. R.

In: Journal of Neuroscience Research, Vol. 27, No. 4, 1990, p. 620-632.

Research output: Contribution to journalArticle

Shan, D. E. ; Beck, C. E. ; Werrbach-Perez, K. ; Perez-Polo, J. R. / Reverse-phase high-performance liquid chromatography of nerve growth factor receptor-like proteins identified with monoclonal antibodies. In: Journal of Neuroscience Research. 1990 ; Vol. 27, No. 4. pp. 620-632.
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