The matrix metalloproteinase gelatinase A in human dentine
The matrix metalloproteinase gelatinase A in human dentine
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DOI：
10.1016/s0003-9969(00)00052-2
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发表时间：
2000-09-01
影响因子：
3
通讯作者：
Overall, CM
中科院分区：
文献类型：
--
作者：
Martin-De las Heras, S;Valenzuela, A;Overall, CM
修改了牙本质蛋白提取方案，以确定基质金属蛋白酶明胶分解活性的非矿化和矿化阶段的人牙本质。依次提取来自15-73岁患者的24个个体恒磨牙的牙本质蛋白，首先用氯化胍（G1提取物），然后用EDTA（E提取物），并且在该脱矿步骤之后，再次用氯化胍（G2提取物）以解离胶原相关蛋白。提取物进行了分析，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和凝胶进行处理，通过蛋白质印迹和酶谱检测明胶分解活性。活性和潜在形式的明胶酶A被确定在非矿化牙本质部分（G1提取物）的58%的牙齿。其他明胶分解物种也被检测到的酶谱与表观M-r的92，54和30 kDa。尽管在所有年龄段的牙齿G1提取物中检测到明胶酶A，表明前牙本质的合成和重塑更近，但在任何E提取物或41岁以上患者的G2提取物中从未检测到明胶酶A。明胶酶A的活性形式在矿化的人牙本质中的存在暗示这种酶在牙本质矿化中。(C)2000爱思唯尔科技有限公司版权所有。
A dentine protein extraction protocol was modified in order to identify matrix metalloproteinase gelatinolytic activities in the non-mineralized and mineralized phases of human dentine. Dentine proteins from 24 individual permanent molars from patients aged 15-73 years were sequentially extracted, first with guanidinium chloride (G1 extract), then EDTA (E extract), and after this demineralization step, again by guanidinium chloride (G2 extract) to dissociate collagen-associated proteins. Extracts were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and the gels were processed by Western blotting and zymography to detect gelatinolytic activities. Active and latent forms of gelatinase A were identified in the non-mineralized dentine fraction (G1 extract) of 58% of the teeth. Other gelatinolytic species were also detected by zymography with apparent M-r of 92, 54 and 30 kDa. Although gelatinase A was detected in the G1 extracts of teeth from all ages, indicating more recent synthesis and remodelling of the predentine, gelatinase A was never detected in any E extract or in the G2 extracts of patients older than 41 years. The presence of the active form of gelatinase A in mineralized human dentine implicates this enzyme in dentine mineralization. (C) 2000 Elsevier Science Ltd. All rights reserved.