This Article Full Text (PDF) References Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Ryu, O.H. Articles by Simmer, J.P. Search for Related Content PubMed PubMed Citation Articles by Ryu, O.H. Articles by Simmer, J.P. Social Bookmarking                         What's this?

Characterization of Recombinant Pig Enamelysin Activity and Cleavage of Recombinant Pig and Mouse Amelogenins

University of Texas Health Science Center at San Antonio, School of Dentistry, Department of Pediatric Dentistry, 7703 Floyd Curl Drive, San Antonio, Texas 78284-7888

A.G. Fincham

Center for Craniofacial Molecular Biology, University of Southern California, School of Dentistry, 2250 Alcazar Street, CSA 1st Floor, Los Angeles, California 90033

C.-C. Hu

University of Texas Health Science Center at San Antonio, School of Dentistry, Department of Pediatric Dentistry, 7703 Floyd Curl Drive, San Antonio, Texas 78284-7888

C. Zhang

University of Texas Health Science Center at San Antonio, School of Dentistry, Department of Pediatric Dentistry, 7703 Floyd Curl Drive, San Antonio, Texas 78284-7888

Q. Qian

University of Texas Health Science Center at San Antonio, School of Dentistry, Department of Pediatric Dentistry, 7703 Floyd Curl Drive, San Antonio, Texas 78284-7888

J.D. Bartlett

Department of Biomineralization, Forsyth Dental Center, 140 Fenway, Boston, Massachusetts 02115

J.P. Simmer

University of Texas Health Science Center at San Antonio, School of Dentistry, Department of Pediatric Dentistry, 7703 Floyd Curl Drive, San Antonio, Texas 78284-7888

Enamelysin (MMP-20) is a tooth-specific matrix metalloproteinase that is initially expressed by ameloblasts and odontoblasts immediately prior to the onset of dentin mineralization, and continues to be expressed throughout the secretory stage of amelogenesis. During the secretory stage, enamel proteins are secreted and rapidly cleaved into a large number of relatively stable cleavage products. Multiple proteinases are present in the developing enamel matrix, and the precise role of enamelysin in the processing of enamel proteins is unknown. We have expressed, activated, and purified the catalytic domain of recombinant pig enamelysin, and expressed a recombinant form of the major secreted pig amelogenin rP172. These proteins were incubated together, and the digestion products were analyzed by SDS-PAGE and mass spectrometric analyses. We assigned amelogenin cleavage products by selecting among the possible polypeptides having a mass within 2 Daltons of the measured values. The polypeptides identified included the intact protein (amino acids 2-173), as well as 2-148, 2-136, 2-107, 2-105, 2-63, 2-45, 46-148, 46-147, 46-107, 46-105, 64-148, 64-147, and 64-136. These fragments of rP172 include virtually all of the major amelogenin cleavage products observed in vivo. We propose that enamelysin is the predominant proteinase that processes enamel proteins during the secretory phase of amelogenesis.

Key Words: enamel • matrix • metalloproteinase • enamelysin • amelogenin • MMP-20

Journal of Dental Research, Vol. 78, No. 3, 743-750 (1999)
DOI: 10.1177/00220345990780030601


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				T. Nagano, A. Kakegawa, Y. Yamakoshi, S. Tsuchiya, J.C.-C. Hu, K. Gomi, T. Arai, J.D. Bartlett, and J.P. Simmer Mmp-20 and Klk4 Cleavage Site Preferences for Amelogenin Sequences Journal of Dental Research, September 1, 2009; 88(9): 823 - 828. [Abstract] [Full Text] [PDF]

				J. P. Simmer, Y. Hu, R. Lertlam, Y. Yamakoshi, and J. C.-C. Hu Hypomaturation Enamel Defects in Klk4 Knockout/LacZ Knockin Mice J. Biol. Chem., July 10, 2009; 284(28): 19110 - 19121. [Abstract] [Full Text] [PDF]

				S.-Y. Kwak, F. B. Wiedemann-Bidlack, E. Beniash, Y. Yamakoshi, J. P. Simmer, A. Litman, and H. C. Margolis Role of 20-kDa Amelogenin (P148) Phosphorylation in Calcium Phosphate Formation in Vitro J. Biol. Chem., July 10, 2009; 284(28): 18972 - 18979. [Abstract] [Full Text] [PDF]

				J. C.-C. Hu, Y. Hu, C. E. Smith, M. D. McKee, J. T. Wright, Y. Yamakoshi, P. Papagerakis, G. K. Hunter, J. Q. Feng, F. Yamakoshi, et al. Enamel Defects and Ameloblast-specific Expression in Enam Knock-out/lacZ Knock-in Mice J. Biol. Chem., April 18, 2008; 283(16): 10858 - 10871. [Abstract] [Full Text] [PDF]

				Y. Yamakoshi, J. C-C. Hu, T. Iwata, K. Kobayashi, M. Fukae, and J. P. Simmer Dentin Sialophosphoprotein Is Processed by MMP-2 and MMP-20 in Vitro and in Vivo J. Biol. Chem., December 15, 2006; 281(50): 38235 - 38243. [Abstract] [Full Text] [PDF]

				H. Nagase, R. Visse, and G. Murphy Structure and function of matrix metalloproteinases and TIMPs Cardiovasc Res, February 15, 2006; 69(3): 562 - 573. [Abstract] [Full Text] [PDF]

				H. Masuya, K. Shimizu, H. Sezutsu, Y. Sakuraba, J. Nagano, A. Shimizu, N. Fujimoto, A. Kawai, I. Miura, H. Kaneda, et al. Enamelin (Enam) is essential for amelogenesis: ENU-induced mouse mutants as models for different clinical subtypes of human amelogenesis imperfecta (AI) Hum. Mol. Genet., March 1, 2005; 14(5): 575 - 583. [Abstract] [Full Text] [PDF]

				J-W Kim, J P Simmer, T C Hart, P S Hart, M D Ramaswami, J D Bartlett, and J C-C Hu MMP-20 mutation in autosomal recessive pigmented hypomaturation amelogenesis imperfecta J. Med. Genet., March 1, 2005; 42(3): 271 - 275. [Full Text] [PDF]

				P S Hart, T C Hart, M D Michalec, O H Ryu, D Simmons, S Hong, and J T Wright Mutation in kallikrein 4 causes autosomal recessive hypomaturation amelogenesis imperfecta J. Med. Genet., July 1, 2004; 41(7): 545 - 549. [Full Text] [PDF]

				J.C.-C. Hu and Y. Yamakoshi ENAMELIN AND AUTOSOMAL-DOMINANT AMELOGENESIS IMPERFECTA Critical Reviews in Oral Biology & Medicine, November 1, 2003; 14(6): 387 - 398. [Abstract] [Full Text] [PDF]

				J. J. Caterina, Z. Skobe, J. Shi, Y. Ding, J. P. Simmer, H. Birkedal-Hansen, and J. D. Bartlett Enamelysin (Matrix Metalloproteinase 20)-deficient Mice Display an Amelogenesis Imperfecta Phenotype J. Biol. Chem., December 13, 2002; 277(51): 49598 - 49604. [Abstract] [Full Text] [PDF]

				T. Aoba and O. Fejerskov DENTAL FLUOROSIS: CHEMISTRY AND BIOLOGY Critical Reviews in Oral Biology & Medicine, March 1, 2002; 13(2): 155 - 170. [Abstract] [Full Text] [PDF]

				C. Qin, R. G. Cook, R. S. Orkiszewski, and W. T. Butler Identification and Characterization of the Carboxyl-terminal Region of Rat Dentin Sialoprotein J. Biol. Chem., January 5, 2001; 276(2): 904 - 909. [Abstract] [Full Text] [PDF]

				R. M. H. Ravindranath, W.-Y. Tam, P. Nguyen, and A. G. Fincham The Enamel Protein Amelogenin Binds to the N-Acetyl-D-glucosamine-mimicking Peptide Motif of Cytokeratins J. Biol. Chem., December 8, 2000; 275(50): 39654 - 39661. [Abstract] [Full Text] [PDF]