Mapping and regulation of the tumor-associated epitope recognized by monoclonal antibody RS-11

Hiroshi Eto, Sam S. Yoon, Barrie P. Bode, Sadao Kamidono, Keishi Makino, Hideyuki Saya, Hideo Nakamura, Kenneth K. Tanabe

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

We have previously described a rat monoclonal antibody, RS-11, which recognizes a tumor-associated antigen common to several species. In the present study, we have cloned and characterized the antigen recognized by RS-11. We screened a phage expression library prepared from HeLa cDNA and identified a clone that reacts with RS-11. DNA sequence analysis revealed that this clone contains sequences of keratin 18 (nucleotides 568-1196). We constructed several glutathione S-transferase fusion proteins and synthetic peptides based on this DNA sequence analysis and examined their reactivity with RS-11 to accurately map the RS-11 epitope. We determined that the epitope resides within a region of seven amino acids on the α-helix 2B domain of keratin 18 in which two amino acids (Leu366 and Lys370) are completely conserved among intermediate filaments as well as other keratin members that are immunoreactive with RS-11. These two residues are sequentially discontinuous but spatially adjacent. The RS-11 epitope is constitutively present in human primary cultured hepatocytes; however, its immunoreactivity with RS-11 is up-regulated by malignant transformation or stimulation with either epidermal growth factor or transforming growth factor alpha.

Original languageEnglish
Pages (from-to)27075-27083
Number of pages9
JournalJournal of Biological Chemistry
Volume275
Issue number35
DOIs
Publication statusPublished - 2000 Sep 1
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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