MMP20 and KLK4 activation and inactivation interactions in vitro.
MMP20 and KLK4 activation and inactivation interactions in vitro.
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DOI：
10.1016/j.archoralbio.2013.08.005
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发表时间：
2013-11
影响因子：
3
通讯作者：
Oida, Shinichiro
中科院分区：
文献类型：
--
作者：
Yamakoshi, Yasuo;Simmer, James P.;Bartlett, John D.;Karakida, Takeo;Oida, Shinichiro
釉质溶解素（MMP 20）和激肽释放酶4（KLK 4）被认为是从发育牙齿的釉质基质中清除蛋白质所必需的。MMP 20由分泌期成釉细胞表达，而KLK 4从过渡期开始在整个成熟期表达。本研究的目的是研究MMP 20对KLK 4的激活和KLK 4对MMP 20的失活。天然猪MMP 20（pMMP 20）和KLK 4（pKLK 4）直接从来自发育磨牙的釉质刮屑中分离。通过与pMMP 20或重组人MMP 20（rhMMP 20）孵育来活化重组人proKLK 4（rh-proKLK 4），并且通过酶谱法检测所得KLK 4活性。通过反相高效液相色谱法（RP-HPLC）分离反应产物，并通过Edman降解表征其N-末端。将pMMP 20与pKLK 4在弱酸性或生理条件下孵育，并通过酶谱法分析酶活性。将rhMMP 20的催化结构域与pKLK 4或重组人KLK 4（rhKLK 4）孵育，并通过酶谱和Edman降解来表征消化产物。pMMP 20和rhMMP 20均通过在体内使用的前肽-酶连接处裂解来活化rh-proKLK 4。pMMP 20在生理条件下被pKLK 4灭活，但在弱酸性条件下不被灭活。pKLK 4和rhKLK 4主要在MMP 20催化结构域的两个位点切割MMP 20。MMP 20在体外激活proKLK 4，KLK 4使MMP 20失活，这些作用可能在体内釉质形成过程中发生。
Enamelysin (MMP20) and kallikrein 4 (KLK4) are believed to be necessary to clear proteins from the enamel matrix of developing teeth. MMP20 is expressed by secretory stage ameloblasts, while KLK4 is expressed from the transition stage throughout the maturation stage. The aim of this study is to investigate the activation of KLK4 by MMP20 and the inactivation of MMP20 by KLK4. Native pig MMP20 (pMMP20) and KLK4 (pKLK4) were isolated directly from enamel scrapings from developing molars. Recombinant human proKLK4 (rh-proKLK4) was activated by incubation with pMMP20 or recombinant human MMP20 (rhMMP20), and the resulting KLK4 activity was detected by zymography. Reaction products were isolated by reverse-phase high performance liquid chromatography (RP-HPLC), and their N-termini characterized by Edman degradation. The pMMP20 was incubated with pKLK4 under mildly acidic or under physiologic conditions, and enzyme activity was analyzed by zymography. The catalytic domain of rhMMP20 was incubated with pKLK4 or recombinant human KLK4 (rhKLK4) and the digestion products were characterized by zymography and Edman degradation. Both pMMP20 and rhMMP20 activated rh-proKLK4 by cleaving at the propeptide-enzyme junction used in vivo. The pMMP20 was inactivated by pKLK4 under physiologic conditions, but not under mildly acidic conditions. Both pKLK4 and rhKLK4 cleaved MMP20 principally at two sites in the catalytic domain of MMP20. MMP20 activates proKLK4 and KLK4 inactivates MMP20 in vitro, and these actions are likely to occur during enamel formation in vivo.
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影响因子：
--
作者：
SMITH, CE;POMPURA, JR;NANCI, A
通讯作者：
NANCI, A
影响因子：
7.6
作者：
Lacruz, R. S.;Smith, C. E.;Paine, M. L.
通讯作者：
Paine, M. L.
影响因子：
7.6
作者：
Chun YH;Yamakoshi Y;Yamakoshi F;Fukae M;Hu JC;Bartlett JD;Simmer JP
通讯作者：
Simmer JP
DOI：
10.1016/s0022-5320(62)80036-7
发表时间：
1962-01-01
期刊：
JOURNAL OF ULTRASTRUCTURE RESEARCH
影响因子：
--
作者：
RONNHOLM, E
通讯作者：
RONNHOLM, E
影响因子：
3.5
作者：
Arendt, Yvonne;Banci, Lucia;Gonnelli, Leonardo
通讯作者：
Gonnelli, Leonardo