Dipeptidyl-peptidase II and cathepsin B activities in amelogenesis of the rat incisor.
A body of published evidence suggests that a significant portion of enamel matrix protein synthesized by ameloblasts localises in the lysosomal-endosomal organelles of these enamel organ cells. Little is known regarding the lysosomal proteolytic activities during amelogenesis. The aims of this study were to detect and measure the activities of lysosomal peptidases cathepsin B (E.C. 3.4.22.1) and dipeptidyl-peptidase II (E.C. 3.4.14.2) in the enamel organ of the rat incisor and to ascertain whether rat enamel matrix proteins are degraded by these peptidases in vitro. Whole enamel organs were dissected from rat mandibular incisors. Enamel protein was also collected from the rat teeth. Analysis indicated that the rat incisor enamel organs contained specific activities of both dipeptidyl-peptidase II and cathepsin B at levels comparable with those of kidney which is rich in both these lysosomal peptidases. Gel electrophoresis and immunoblotting demonstrated that both cathepsin B and dipeptidyl-peptidase II were able to substantially degrade the rat enamel proteins in vitro. Based on these observations, we propose that lysosomal proteases have roles in amelogenesis in the intracellular degradation of amelogenins.
Smid
J R
JR
Oral Biology and Pathology, School of Dentistry, The University of Queensland, Brisbane, Australia. j.smid@mailbox.uq.edu.au
Young
W G
WG
Monsour
P A
PA
D000566:Amelogenesis / Q000502:physiology*; D053523:Amelogenin; D000818:Animals; D015153:Blotting, Western; D002401:Cathepsin B / Q000032:analysis / Q000378:metabolism*; D002863:Chromogenic Compounds; D003374:Coumarins; D003746:Dental Enamel Proteins / Q000032:analysis / Q000378:metabolism; D004151:Dipeptides; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000032:analysis / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004658:Enamel Organ / Q000201:enzymology*; D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D007180:Incisor; D007668:Kidney / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology; D051381:Rats; D017208:Rats, Wistar; D013050:Spectrometry, Fluorescence; D013223:Statistics as Topic
D053523:Amelogenin; C507201:Amelx protein, rat; D002863:Chromogenic Compounds; D003374:Coumarins; D003746:Dental Enamel Proteins; D004151:Dipeptides; D005456:Fluorescent Dyes; C109069:enamel matrix proteins; C069012:benzyloxycarbonylarginyl-arginine 4-methylcoumarin-7-ylamide; C045792:7-lysylalanyl-4-methylcoumarinamide; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; C100151:dipeptidyl peptidase II; D002401:Cathepsin B