title: DPPI may activate KLK4 during enamel formation.
abstract: Kallikrein-4 (KLK4) is a serine protease expressed during enamel maturation, and proteolytic processing of the enamel matrix by KLK4 is critical for proper enamel formation. KLK4 is secreted as an inactive zymogen (pro-KLK4), and identification of its activator remains elusive. Dipeptidyl peptidase I (DPPI) is a cysteine aminopeptidase that can activate several serine proteases. In this study, we sought to examine DPPI expression in mouse enamel organ and determine if DPPI could activate KLK4. Real-time PCR showed DPPI expression throughout amelogenesis, with highest expression at maturation, and immunohistochemical staining of mouse incisors confirmed DPPI expression by ameloblasts. We demonstrate in vitro that DPPI activates pro-KLK4 to cleave a fluorogenic peptide containing a KLK4 cleavage site. Examination of mature enamel from DPPI null mice by FTIR showed no significant accumulation of protein; however, microhardness testing revealed that loss of DPPI expression significantly reduced enamel hardness.
authors.affiliation: Department of Cytokine Biology, Forsyth Institute, Harvard School of Dental Medicine, 140 The Fenway, Boston, MA 02115, USA.
authors.forename: C E
authors.initials: CE
authors.lastname: Tye
authors.forename: C T
authors.initials: CT
authors.lastname: Pham
authors.forename: J P
authors.initials: JP
authors.lastname: Simmer
authors.forename: J D
authors.initials: JD
authors.lastname: Bartlett
mesh_terms: D000566:Amelogenesis / Q000235:genetics / Q000502:physiology*; D000818:Animals; D020789:Cathepsin C / Q000235:genetics / Q000378:metabolism*; D003743:Dental Enamel / Q000201:enzymology* / Q000648:ultrastructure; D003746:Dental Enamel Proteins / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D007610:Kallikreins / Q000235:genetics / Q000378:metabolism*; D008334:Mandible; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D018345:Mice, Knockout; D008817:Mice, Mutant Strains; D008963:Molar / Q000648:ultrastructure; D011994:Recombinant Proteins; D013045:Species Specificity; D014074:Tooth Calcification / Q000235:genetics / Q000502:physiology*
chemical_list: D003746:Dental Enamel Proteins; D011994:Recombinant Proteins; D020789:Cathepsin C; C533919:Ctsc protein, mouse; D007610:Kallikreins; C120801:kallikrein 4