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Identification of Full-Length Dentin Matrix Protein 1 in Dentin and Bone

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Abstract

Dentin matrix protein 1 (DMP1) has been identified in the extracellular matrix (ECM) of dentin and bone as the processed NH2-terminal and COOH-terminal fragment. However, the full-length form of DMP1 has not been identified in these tissues. The focus of this investigation was to search for the intact full-length DMP1 in dentin and bone. We used two types of anti-DMP1 antibodies to identify DMP1: one type specifically recognizes the NH2-terminal region and the other type is only reactive to the COOH-terminal region of the DMP1 amino acid sequence. An ∼105-kDa protein, extracted from the ECM of rat dentin and bone, was recognized by both types of antibodies; and the migration rate of this protein was identical to the recombinant mouse full-length DMP1 made in eukaryotic cells. We concluded that this ∼105-kDa protein is the full-length form of DMP1, which is considerably less abundant than its processed fragments in the ECM of dentin and bone. We also detected the full-length form of DMP1 and its processed fragments in the extract of dental pulp/odontoblast complex dissected from rat teeth. In addition, immunofluorescence analysis showed that in MC3T3-E1 cells the NH2-terminal and COOH-terminal fragments of DMP1 are distributed differently. Our findings indicate that the majority of DMP1 must be cleaved within the cells that synthesize it and that minor amounts of uncleaved DMP1 molecules are secreted into the ECM of dentin and bone.

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References

  1. George A, Sabsay B, Simonian PAL, Veis A (1993) Characterization of a novel dentin matrix acidic phosphoprotein: implications for induction of biomineralization. J Biol Chem 268:12624–12630

    PubMed  CAS  Google Scholar 

  2. Hirst KL, Ibaraki-O’Connor K, Young MF, Dixon MJ (1997) Cloning and expression analysis of the bovine dentin matrix acidic phosphoprotein gene. J Dent Res 76:754–760

    Article  PubMed  CAS  Google Scholar 

  3. MacDougall M, Gu T, Luan X, Simmons D, Chen J (1998) Identification of a novel isoform of mouse dentin matrix protein 1: spatial expression in mineralized tissues. J Bone Miner Res 13:422–431

    Article  PubMed  CAS  Google Scholar 

  4. Narayanan K, Srinivas R, Ramachandran A, Hao J, Quinn B, George A (2001) Differentiation of embryonic mesenchymal cells to odontoblast-like cells by overexpression of dentin matrix protein 1. Proc Natl Acad Sci USA 98:4516–4521

    Article  PubMed  CAS  Google Scholar 

  5. Ye L, MacDougall M, Zhang S, Xie Y, Zhang J, Li Z, Lu Y, Mishina Y, Feng JQ (2004) Deletion of dentin matrix protein-1 leads to a partial failure of maturation of predentin into dentin, hypomineralization, and expanded cavities of pulp and root canal during postnatal tooth development. J Biol Chem 279:19141–19148

    Article  PubMed  CAS  Google Scholar 

  6. Ye L, Mishina Y, Chen D, Huang H, Dallas SL, Dallas MR, Sivakumar P, Kunieda T, Tsutsui TW, Boskey A, Bonewald LF, Feng JQ (2005) Dmp1-deficient mice display severe defects in cartilage formation responsible for a chondrodysplasia-like phenotype. J Biol Chem 280:6197–6203

    Article  PubMed  CAS  Google Scholar 

  7. Feng JQ, Ward LM, Liu S, Lu Y, Xie Y, Yuan B, Yu X, Rauch F, Davis SI, Zhang S, Rios H, Drezner MK, Quarles LD, Bonewald LF, White KE (2006) Loss of DMP1 causes rickets and osteomalacia and identifies a role for osteocytes in mineral metabolism. Nat Genet 38:1230–1235

    Article  CAS  Google Scholar 

  8. Narayanan K, Ramachandran A, Hao J, He G, Park KW, Cho M, George A (2003) Dual functional roles of dentin matrix protein 1. Implications in biomineralization and gene transcription by activation of intracellular Ca2+ store. J Biol Chem 278:17500–17508

    Article  PubMed  CAS  Google Scholar 

  9. Lu Y, Ye L, Yu S, Zhang S, Xie Y, McKee MD, Li YC, Kong J, Eick JD, Dallas SL, Feng JQ (2007) Rescue of odontogenesis in Dmp1-deficient mice by targeted re-expression of DMP1 reveals roles for DMP1 in early odontogenesis and dentin apposition in vivo. Dev Biol 303:191–201

    Article  PubMed  CAS  Google Scholar 

  10. Toyosawa S, Sato A, O’hUigin C, Tichy H, Klein J (2000) Expression of the dentin matrix protein 1 gene in birds. J Mol Evol 50:31–38

    PubMed  CAS  Google Scholar 

  11. Kim JW, Yamakoshi Y, Iwata T, Hu YY, Zhang H, Hu JC, Simmer JP (2006) Porcine dentin matrix protein 1: gene structure, cDNA sequence, and expression in teeth. Eur J Oral Sci 1University of Michigan Dental Research Laboratory, Ann Arbor, MI, USA 114:33–41

    CAS  Google Scholar 

  12. Qin C, Brunn JC, Cook RG, Orkiszewski RS, Malone JP, Veis A, Butler WT (2003) Evidence for the proteolytic processing of dentin matrix protein 1. Identification and characterization of processed fragments and cleavage sites. J Biol Chem 278:34700–34708

    Article  PubMed  CAS  Google Scholar 

  13. Qin C, Huang B, Wygant JN, McIntyre BW, McDonald CH, Cook RG, Butler WT (2006) A chondroitin sulfate chain attached to the bone dentin matrix protein 1 NH2-terminal fragment. J Biol Chem 281:8034–8040

    Article  PubMed  CAS  Google Scholar 

  14. Tartaix PH, Doulaverakis M, George A, Fisher LW, Butler WT, Qin C, Salih E, Tan M, Fujimoto Y, Spevak L, Boskey AL (2004) In vitro effects of dentin matrix protein-1 on hydroxyapatite formation provide insights into in vivo functions. J Biol Chem 279:18115–18120

    Article  PubMed  CAS  Google Scholar 

  15. He G, Gajjeraman S, Schultz D, Cookson D, Qin C, Butler WT, Hao J, George A (2005) Spatially and temporally controlled biomineralization is facilitated by interaction between self-assembled dentin matrix protein 1 and calcium phosphate nuclei in solution. Biochemistry 44:16140–16148

    Article  PubMed  CAS  Google Scholar 

  16. Gajjeraman S, Narayanan K, Hao J, Qin C, George A (2007) Matrix macromolecules in hard tissues control the nucleation and hierarchical assembly of hydroxyapatite. J Biol Chem 282:1193–1204

    Article  PubMed  CAS  Google Scholar 

  17. Butler WT, Bhown M, Dimuzio MT, Linde A (1981) Nonocollagenous proteins of dentin: isolation and partial characterization of rat dentin proteins and proteoglycans using a three-step preparative method. Coll Relat Res 1:187–199

    PubMed  CAS  Google Scholar 

  18. Qin C, Brunn JC, Jones J, George A, Ramachandran A, Gorski JP, Butler WT (2001) A comparative study of sialic acid-rich proteins in rat bone and dentin. Eur J Oral Sci 109:133–141

    Article  PubMed  CAS  Google Scholar 

  19. Qin C, Baba O, Brunn JC, McKee MD, Bonewald L, Butler WT (2005) Dentin matrix protein 1 (DMP1) and dentin sialophosphoprotein (DSPP) share unique properties including tissue localization, proteolytic processing, and high molecular weight forms. In: Sodek J, Landis W (eds) Proceedings of the 8th ICCBMT. University of Toronto Press, Toronto, pp 174–177

  20. Baba O, Qin C, Brunn JC, Wygant JN, McIntyre BW, Butler WT (2004) Colocalization of dentin matrix protein 1 and dentin sialoprotein at late stages of rat molar development. Matrix Biol 23:371–379

    Article  PubMed  CAS  Google Scholar 

  21. Fisher LW, Torchia DA, Fohr B, Young MF, Fedarko NS (2001) Flexible structures of SIBLING proteins, bone sialoprotein, and osteopontin. Biochem Biophys Res Commun 280:460–465

    Article  PubMed  CAS  Google Scholar 

  22. Narayanan K, Gajjeraman S, Ramachandran A, Hao J, George A (2006) Dentin matrix protein 1 regulates dentin sialophosphoprotein gene transcription during early odontoblast differentiation. J Biol Chem 281:19064–19071

    Article  PubMed  CAS  Google Scholar 

  23. Takahara K, Lyons GE, Greenspan DS (1994) Bone morphogenetic protein-1 and a mammalian tolloid homologue (mTld) are encoded by alternatively spliced transcripts which are differentially expressed in some tissues. J Biol Chem 269:32572–32578

    PubMed  CAS  Google Scholar 

  24. Steiglitz BM, Ayala M, Narayanan K, George A, Greenspan DS (2004) Bone morphogenetic protein-1/Tolloid-like proteinases process dentin matrix protein-1. J Biol Chem 279:980–986

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This work was supported by National Institutes of Health grant DE 005092 (to C. Q.). I. M. is on sabbatical leave from the Medical University of Gdansk, Poland.

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Authors and Affiliations

  1. Department of Biomedical Sciences, Baylor College of Dentistry, Texas A & M University System Health Science Center, 3302 Gaston Avenue, Dallas, TX, 75246, USA

    Bingzhen Huang, Izabela Maciejewska, Yao Sun, Tao Peng, Disheng Qin, William T. Butler, Jian Feng & Chunlin Qin

  2. Department of Oral Biology, University of Missouri at Kansas City, 650 E. 25th St., Kansas City, MO, 64108, USA

    Yongbo Lu & Lynda Bonewald

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  1. Bingzhen Huang
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  2. Izabela Maciejewska
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  3. Yao Sun
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  4. Tao Peng
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  5. Disheng Qin
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  6. Yongbo Lu
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  8. William T. Butler
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  9. Jian Feng
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  10. Chunlin Qin
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Correspondence to Chunlin Qin.

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Huang, B., Maciejewska, I., Sun, Y. et al. Identification of Full-Length Dentin Matrix Protein 1 in Dentin and Bone. Calcif Tissue Int 82, 401–410 (2008). https://doi.org/10.1007/s00223-008-9140-7

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  • DOI: https://doi.org/10.1007/s00223-008-9140-7

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