Histochemistry and Cell Biology

, Volume 107, Issue 6, pp 485–494

Immunochemical and immunohistochemical study of the 27- and 29-kDa calcium-binding proteins and related proteins in the porcine tooth germ

  • C. Murakami
  • N. Dohi
  • M. Fukae
  • T. Tanabe
  • Y. Yamakoshi
  • K. Wakida
  • T. Satoda
  • O. Takahashi
  • M. Shimizu
  • O. H. Ryu
  • J. P. Simmer
  • T. Uchida
ORIGINAL PAPER

DOI: 10.1007/s004180050136

Cite this article as:
Murakami, C., Dohi, N., Fukae, M. et al. Histochemistry (1997) 107: 485. doi:10.1007/s004180050136

Abstract

 Our previous report identified 27- and 29-kDa calcium-binding proteins in porcine immature dental enamel. In this study we revealed that the N-terminal amino acid sequences of the two proteins were identical: LLANPXGXIPNLARGPAGRSRGPPG. The sequence matches a portion of the amino acid sequence of the porcine sheath protein, sheathlin. Porcine tooth germs were investigated immunochemically and immunohistochemically using specific antibodies raised against synthetic peptide that included residues 13–25 of this sequence. The affinity-purified antibodies reacted with several proteins extracted from newly formed immature enamel in immunochemical analyses, especially protein bands migrating at 62, 35–45, 29, and 27 kDa in SDS-polyacrylamide gels. The largest protein detected was a weak band near 70 kDa. In immunochemical analyses of proteins extracted from the inner (old) immature enamel, the antibody reacted faintly with the 27- and 29-kDa proteins. In immunohistochemical preparations, the Golgi apparatus and secretory granules of the secretory ameloblast, and the surface layer of immature enamel showed immunoreactivity. The immunoreactivity of immature enamel just beneath the secretory face of the Tomes’ process was intense. No immunoreactivity was found in the Golgi apparatus of the maturation ameloblast. These results suggest that the 70-kDa protein, whose degradation might be very fast, is the parent protein of the 27- and 29-kDa proteins.

Copyright information

© Springer-Verlag Berlin Heidelberg 1997

Authors and Affiliations

  • C. Murakami
    • 1
  • N. Dohi
    • 2
  • M. Fukae
    • 3
  • T. Tanabe
    • 3
  • Y. Yamakoshi
    • 3
  • K. Wakida
    • 1
  • T. Satoda
    • 1
  • O. Takahashi
    • 1
  • M. Shimizu
    • 3
  • O. H. Ryu
    • 4
  • J. P. Simmer
    • 4
  • T. Uchida
    • 1
  1. 1.Department of Oral Anatomy, Hiroshima University School of Dentistry, Kasumi 1-2-3, Minami-ku, Hiroshima, 734, Japan Tel. +81-82-257-5624; Fax +81-82-257-5689 e-mail chikage@ipc.hiroshima-u.ac.jpJP
  2. 2.Department of Endodontology and Periodontology, Hiroshima University School of Dentistry, Kasumi 1-2-3, Minami-ku, Hiroshima, 734, JapanJP
  3. 3.Department of Biochemistry, School of Dental Medicine, Tsurumi University, Tsurumi 2-1-3, Tsurumi-ku, Yokohama 230, JapanJP
  4. 4.Department of Pediatric Dentistry, School of Dentistry, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78284-7888, USAUS