title: Enamel proteins and proteases in Mmp20 and Klk4 null and double-null mice.
abstract: Matrix metalloproteinase 20 (MMP20) and kallikrein-related peptidase 4 (KLK4) are thought to be necessary to clear proteins from the enamel matrix of developing teeth. We characterized Mmp20 and Klk4 null mice to better understand their roles in matrix degradation and removal. Histological examination showed retained organic matrix in Mmp20, Klk4, and Mmp20/Klk4 double-null mouse enamel matrix, but not in the wild-type. X-gal histostaining of Mmp20 null mice heterozygous for the Klk4 knockout/lacZ knockin showed that Klk4 is expressed normally in the Mmp20 null background. This finding was corroborated by zymogram and western blotting, which discovered a 40-kDa protease induced in the maturation stage of Mmp20 null mice. Proteins were extracted from secretory-stage or maturation-stage maxillary first molars from wild-type, Mmp20 null, Klk4 null, and Mmp20/Klk4 double-null mice and were analyzed by SDS-PAGE and western blotting. Only intact amelogenins and ameloblastin were observed in secretory-stage enamel of Mmp20 null mice, whereas the secretory-stage matrix from Klk4 null mice was identical to the matrix from wild-type mice. More residual matrix was observed in the double-null mice compared with either of the single-null mice. These results support the importance of MMP20 during the secretory stage and of KLK4 during the maturation stage and show there is only limited functional redundancy for these enzymes.
authors.affiliation: Department of Biologic and Materials Sciences, University of Michigan School of Dentistry, Ann Arbor, MI 48108, USA.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Yamakoshi
authors.forename: Amelia S
authors.initials: AS
authors.lastname: Richardson
authors.forename: Stephanie M
authors.initials: SM
authors.lastname: Nunez
authors.forename: Fumiko
authors.initials: F
authors.lastname: Yamakoshi
authors.forename: Rachel N
authors.initials: RN
authors.lastname: Milkovich
authors.forename: Jan C-C
authors.initials: JC
authors.lastname: Hu
authors.forename: John D
authors.initials: JD
authors.lastname: Bartlett
authors.forename: James P
authors.initials: JP
authors.lastname: Simmer
mesh_terms: D000565:Ameloblasts / Q000201:enzymology; D000566:Amelogenesis / Q000235:genetics / Q000502:physiology*; D053523:Amelogenin / Q000378:metabolism; D000818:Animals; D003743:Dental Enamel / Q000201:enzymology*; D003746:Dental Enamel Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism; D055786:Gene Knockout Techniques; D005838:Genotype; D007610:Kallikreins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D053519:Matrix Metalloproteinase 20 / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010447:Peptide Hydrolases / Q000378:metabolism; D059748:Proteolysis
chemical_list: C490890:Ambn protein, mouse; D053523:Amelogenin; D003746:Dental Enamel Proteins; D010447:Peptide Hydrolases; D007610:Kallikreins; C120801:kallikrein 4; D053519:Matrix Metalloproteinase 20; C507113:Mmp20 protein, mouse