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Journal of Biochemistry Advance Access originally published online on July 31, 2008
Journal of Biochemistry 2008 144(4):531-537; doi:10.1093/jb/mvn096
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© 2008 The Japanese Biochemical Society.

The Effects of LAMP1 and LAMP3 on M180 Amelogenin Uptake, Localization and Amelogenin mRNA Induction by Amelogenin Protein

Liming Xu1, Hidemitsu Harada2 and Akiyoshi Taniguchi1,*

1Advanced Medical Materials Group, Biomaterials Center, National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044; and 2Department of Oral Anatomy II, Iwate Medical University, School of Dentistry, 1-3-27 Chuodori, Morioka, Iwate 020-8505, Japan

*To whom correspondence should be addressed. Tel: +81-29-860-4505, Fax: +81-29-860-4714, E-mail: taniguchi.akiyoshi{at}nims.go.jp

Received June 26, 2008; Accepted July 22, 2008


  Abstract

We previously demonstrated that the uptake of M180 amelogenin protein in dental epithelial cells (HAT-7) results in increased levels of amelogenin mRNA through enhanced mRNA stabilization. To determine the processes involved in the uptake of extracellular M180 amelogenin by cells and in amelogenin intracellular trafficking in the amelogenin protein-mediated amelogenin mRNA expression pathway, we investigated the effects of LAMP1 and LAMP3, which are candidate M180 amelogenin receptors, on M180 amelogenin uptake, localization and amelogenin mRNA induction by amelogenin protein, using anti-LAMP-1 and anti-LAMP-3 antibodies and siRNA analysis. The results indicate that LAMP3 blocking by anti-LAMP-3 decreases M180 amelogenin uptake, but does not affect amelogenin mRNA induction by amelogenin protein, suggesting that LAMP3 is related to amelogenin degradation. Down-regulation by siRNA of LAMP1, which is the receptor for small amelogenin protein (LRAP), does not affect M180 amelogenin uptake, localization or amelogenin mRNA induction by amelogenin protein. Thus, while LAMP1 is the specific receptor for LRAP, it is not a receptor for M180 amelogenin. These findings will aid further research into the understanding of M180 amelogenin function and expression.

Key Words: amelogenin, LAMP1, LAMP3, ameloblast, tooth development

Abbreviations: FITC, fluorescein isothiocyanate; LAMP, lysosomal associated membrane protein; LRAP, leucine-rich amelogenin peptides


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