Abstract：
:Enamelysin is a matrix metalloproteinase (MMP-20) secreted by ameloblasts, previously shown to hydrolyze recombinant amelogenin. The purpose of this study was to use recombinant MMP-20 to further investigate the specific hydrolysis of peptide fragments containing cleavage sites for tyrosine-rich amelogenin peptide (TRAP) and leucine-rich amelogenin peptide (LRAP). MMP-20 cDNA was isolated from a subtracted bovine cDNA library, reconstructed into pRSET A vector, and overexpressed in BL21 Escherichia coli. The recombinant MMP-20 was purified using Mono-S ion exchange and nickel affinity chromatography. The proteinase was renatured by dialysis against buffer containing 50 microM zinc and 5 mM calcium and autolysed to form several active fragments. The varying sizes and activities of the activated enzyme fragments appeared to be due to sequential autolysis at different location of the carboxyl terminus of the intact enzyme. Two synthetic peptides corresponding to amelogenin amino acid sequences 36-49 and 181-188 were hydrolyzed by the activated rMMP-20. Mass spectrometry and amino acid composition analysis showed that the cleavage sites were between the tryptophan and leucine (45 and 46) for TRAP and between proline and alanine (186-187) for LRAP. These results indicate that MMP-20 can be autoactivated, and activated MMP-20 has a functional role in the initial cleavage of amelogenin.
journal_name
Eur J Oral Scijournal_title
European journal of oral sciencesauthors
Li W,Machule D,Gao C,DenBesten PKdoi
10.1046/j.0909-8836.1999.eos107506.xsubject
Has Abstractpub_date
1999-10-01 00:00:00pages
352-9issue
5eissn
0909-8836issn
1600-0722journal_volume
107pub_type
杂志文章abstract：:The aim of the present study was to examine to what extent pH affects human fibroblast repopulation of an experimental wound in vitro laying special emphasis on cell migration, DNA synthesis, and alterations in cell morphology. Wounds were simulated in vitro by partially scraping off monolayers of human embryonic dipl...
journal_title：European journal of oral sciences
pub_type： 杂志文章
doi：10.1111/j.1600-0722.1995.tb00016.x
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abstract：:The Enamel Defects Index (EDI) was created based on three innovative principles: (i) a basic level of the three major categories of defects; (ii) more detailed subcategories of each major category; and (iii) each category scored independently as present [1] or absent [0], simplifying decision making. The aim of this i...
journal_title：European journal of oral sciences
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journal_title：European journal of oral sciences
pub_type： 杂志文章
doi：10.1111/j.1600-0722.1995.tb00121.x
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journal_title：European journal of oral sciences
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pub_type： 临床试验,杂志文章,随机对照试验
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pub_type： 杂志文章,随机对照试验
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journal_title：European journal of oral sciences
pub_type： 杂志文章
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journal_title：European journal of oral sciences
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journal_title：European journal of oral sciences
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