Salivary carbonic anhydrase isoenzyme VI is located in the human enamel pellicle
Salivary carbonic anhydrase isoenzyme VI is located in the human enamel pellicle
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DOI：
10.1159/000016515
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发表时间：
1999-05-01
期刊：
影响因子：
4.2
通讯作者：
Rajaniemi, H
中科院分区：
文献类型：
--
作者：
Leinonen, J;Kivelä, J;Rajaniemi, H
唾液碳酸酐酶（CA VI）似乎通过直接调节唾液pH和缓冲能力以外的机制来保护牙齿免受龋齿。为了阐明CA VI是否在牙齿表面的局部微环境中起作用，我们研究了人类釉质表膜中酶的位置和活性。使用特异性兔抗人CA VI血清结合免疫染色和免疫印迹技术进行研究。CA活性用组织化学染色法证明。具有体内形成的表膜的提取的牙齿的CA VI免疫染色显示酶存在于釉质表膜中。已知存在于表膜中的唾液α-淀粉酶的免疫染色显示类似的染色模式。通过对在体内形成的表膜蛋白进行免疫印迹，证实了釉质表膜中CA VI的存在。体外研究表明，CA VI结合抛光釉质表面从唾液和纯化的酶的解决方案。釉质表面CA VI免疫染色的强度取决于所用酶的浓度。体外形成的釉膜的组织化学染色证实，结合酶保留其酶活性。人牙釉质表膜中存在活性CA VI表明其可通过在牙齿表面的表膜层中局部发挥作用来加速酸的去除。
Salivary carbonic anhydrase (CA VI) appears to protect teeth from caries via mechanisms other than direct regulation of salivary pH and buffering capacity. To elucidate whether CA VI acts in the local microenvironment of the tooth surface, we studied the location and activity of the enzyme in the human enamel pellicle. The study was performed using a specific rabbit antiserum to human CA VI in conjunction with immunostaining and immunoblot techniques. CA activity was demonstrated using a histochemical staining method. CA VI immunostaining of extracted teeth having in vivo formed pellicle showed that the enzyme is present in the enamel pellicle, Immunostaining for salivary alpha-amylase, which is known to be present in the pellicle, showed a similar staining pattern. The presence of CA VI in the enamel pellicle was confirmed by immunoblotting of in vivo formed pellicle proteins. In vitro studies showed that CA VI binds to polished enamel surfaces from both saliva and solutions of purified enzyme. The intensity of the CA VI immunostaining on the enamel surface was dependent on the concentration of the applied enzyme. The histochemical staining of in vitro formed enamel pellicle confirmed that the bound enzyme retains its enzymatic activity. The presence of active CA VI in the human enamel pellicle suggests that it may accelerate the removal of acid by functioning locally in the pellicle layer on dental surfaces.