A monoclonal antibody, 3A10, recognizes a specific amino acid sequence present on a series of developmentally expressed brain proteins

Ayako Harada, Ken Ichi Takeuchi, Naoshi Dohmae, Koji Takio, Toshimitu Uenaka, Junken Aoki, Keizo Inoue, Masato Umeda

Research output: Contribution to journalArticlepeer-review

12 Citations (Scopus)

Abstract

Immunoblotting showed that a monoclonal antibody, 3A10, binds to a series of rat brain-specific antigens with molecular masses of 150-, 120-, 118-, 106-, 104-, 79-, and 77-kDa. The expression of 3A10 antigens is dependent on the developmental stage of the brain; only the 106-kDa antigen is detected during embryonic stages of rat brain development, while the expression of the remaining 6 antigens starts after birth and reaches a maximum during postnatal days 15-21. Detection of the 3A10 antigens in cultured neuronal and glial cells derived from cerebral cortices of rat brain at embryonic day 18 showed that the 77-, 79-, 106-, and 150-kDa antigens are specifically expressed in neuronal cells. The 77-kDa antigen was purified and identified as synapsin I by amino acid sequence analyses of the peptide fragments isolated after Achromobacter protease I treatment. During the isolation of 3A10-reactive proteins by immunological screening of cDNA libraries constructed from adult rat brain, we found that all of the 3A10-reactive clones contain nucleotide sequences encoding the unique amino acid sequence TRSP(S,R,G)P. Analyses of 3A10-binding to various synthetic peptides showed that the monoclonal antibody recognizes a specific conformational structure formed by either the TRSPXP sequence or similar amino acid sequences that are expressed on a series of developmentally expressed brain proteins.

Original languageEnglish
Pages (from-to)443-448
Number of pages6
JournalJournal of biochemistry
Volume125
Issue number3
DOIs
Publication statusPublished - 1999

Keywords

  • Brain
  • Development
  • Monoclonal antibody
  • Neuron
  • Synapsin I

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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