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Cleavage site specificity of MMP-20 for secretory-stage ameloblastin
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نویسنده
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chun y.-h.p. ,yamakoshi y. ,yamakoshi f. ,fukae m. ,hu j.c.-c. ,bartlett j.d. ,simmer j.p.
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منبع
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journal of dental research - 2010 - دوره : 89 - شماره : 8 - صفحه:785 -790
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چکیده
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Ameloblastin is processed by protease(s) during enamel formation. we tested the hypothesis that mmp-20 (enamelysin) catalyzes the cleavages that generate secretory-stage ameloblastin cleavage products. we isolated a 23-kda ameloblastin cleavage product from developing enamel and determined its n-terminus sequence. ameloblastin was stably expressed and secreted from hek293-h cells,purified,and digested with mmp-20 or klk4 (kallikrein 4). the digests were analyzed by sds-page and western blotting,and cleavage products were characterized by n-terminal sequencing. six fluorescent peptides were digested with mmp-20 and klk4 and analyzed by rp-hplc and by mass spectrometry. mmp-20 cleaved each peptide exactly at the sites corresponding to ameloblastin cleavages catalyzed in vivo. klk4 cleaved ameloblastin and the fluorescent peptides at sites not observed in vivo,and cleaved at only a single correct site: before leu171. we conclude that mmp-20 is the enzyme that processes ameloblastin during the secretory stage of amelogenesis,and we present a hypothesis about the sequence of ameloblastin cleavages. © international & american associations for dental research.
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کلیدواژه
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enamel; fluorescent peptides; proteases; tooth
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آدرس
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department of biologic and materials sciences,university of michigan,school of dentistry,1011 north university,ann arbor,mi 48109-1078,united states,department of periodontics,university of texas,health science center at san antonio,7703 floyd curl drive,san antonio,tx 78229, United States, department of biologic and materials sciences,university of michigan,school of dentistry,1011 north university,ann arbor,mi 48109-1078, United States, department of biologic and materials sciences,university of michigan,school of dentistry,1011 north university,ann arbor,mi 48109-1078, United States, department of biochemistry,school of dental medicine,tsurumi university,2-1-3 tsurumi,tsurumi-ku,yokohama 230, Japan, department of biologic and materials sciences,university of michigan,school of dentistry,1011 north university,ann arbor,mi 48109-1078, United States, department of cytokine biology,forsyth institute,140 the fenway,boston,ma 02115, United States, department of biologic and materials sciences,university of michigan,school of dentistry,1011 north university,ann arbor,mi 48109-1078, United States
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Authors
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