Discovery of a novel and rich source of gluten-degrading microbial enzymes in the oral cavity.
Discovery of a novel and rich source of gluten-degrading microbial enzymes in the oral cavity.
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DOI：
10.1371/journal.pone.0013264
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发表时间：
2010-10-11
期刊：
影响因子：
3.7
通讯作者：
Oppenheim FG
中科院分区：
文献类型：
--
作者：
Helmerhorst EJ;Zamakhchari M;Schuppan D;Oppenheim FG
乳糜泻是一种T细胞介导的炎症性肠病，由携带HLA-DQ 2或HLA-DQ 8的遗传易感个体摄入麸质引起。含有多个谷氨酰胺和脯氨酸残基的免疫原性麦胶蛋白表位在很大程度上抵抗胃和肠蛋白酶的降解。然而，唾液微生物表现出谷氨酰胺内切蛋白酶活性，发现针对富含谷氨酰胺和脯氨酸的唾液蛋白。目的是探索醇溶蛋白是否可以作为口腔微生物酶的底物。研究了悬浮牙菌斑中的蛋白水解活性，所述蛋白水解活性针对a）麦醇溶蛋白衍生的对硝基苯胺（pNA）连接的合成酶底物B）天然麦醇溶蛋白的混合物和c）合成的高免疫原性麦醇溶蛋白肽（α2-麦醇溶蛋白的33-mer和γ-麦醇溶蛋白的26-mer）。此外，进行麦醇溶蛋白酶谱分析以获得麦醇溶蛋白降解口服酶的近似分子量和pH活性谱，并进行液体等电聚焦以建立总体酶等电点。噬菌斑细菌有效地水解Z-YPQ-pNA、Z-QQP-pNA、Z-PPF-pNA和Z-PFP-pNA，其中Z-YPQ-pNA被最快速地切割。麦醇溶蛋白免疫原性结构域在口腔细菌的存在下被广泛降解。醇溶蛋白酶谱显示，突出的酶表现出分子量>70 kD，并在3至10的宽pH范围内具有活性。液体等电聚焦表明，大多数麦醇溶蛋白降解酶是酸性的，等电点在2.5和4.0之间。这是第一次报道与上胃肠道相关的麸质降解微生物的证据。这些微生物可能在消化膳食麸质中发挥迄今未被认识到的作用，从而保护处于风险中的受试者免受乳糜泻。
Celiac disease is a T cell mediated-inflammatory enteropathy caused by the ingestion of gluten in genetically predisposed individuals carrying HLA-DQ2 or HLA-DQ8. The immunogenic gliadin epitopes, containing multiple glutamine and proline residues, are largely resistant to degradation by gastric and intestinal proteases. Salivary microorganisms however exhibit glutamine endoprotease activity, discovered towards glutamine- and proline-rich salivary proteins. The aim was to explore if gliadins can serve as substrates for oral microbial enzymes. Proteolytic activity in suspended dental plaque was studied towards a) gliadin-derived paranitroanilide(pNA)-linked synthetic enzyme substrates b) a mixture of natural gliadins and c) synthetic highly immunogenic gliadin peptides (33-mer of α2-gliadin and 26-mer of γ-gliadin). In addition, gliadin zymography was conducted to obtain the approximate molecular weights and pH activity profiles of the gliadin-degrading oral enzymes and liquid iso-electric focusing was performed to establish overall enzyme iso-electric points. Plaque bacteria efficiently hydrolyzed Z-YPQ-pNA, Z-QQP-pNA, Z-PPF-pNA and Z-PFP-pNA, with Z-YPQ-pNA being most rapidly cleaved. Gliadin immunogenic domains were extensively degraded in the presence of oral bacteria. Gliadin zymography revealed that prominent enzymes exhibit molecular weights >70 kD and are active over a broad pH range from 3 to 10. Liquid iso-electric focusing indicated that most gliadin-degrading enzymes are acidic in nature with iso-electric points between 2.5 and 4.0. This is the first reported evidence for gluten-degrading microorganisms associated with the upper gastro-intestinal tract. Such microorganisms may play a hitherto unappreciated role in the digestion of dietary gluten and thus protection from celiac disease in subjects at risk.
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影响因子：
7
作者：
Messana, Irene;Cabras, Tiziana;Castagnola, Massimo
通讯作者：
Castagnola, Massimo
影响因子：
29.4
作者：
Camp JG;Kanther M;Semova I;Rawls JF
通讯作者：
Rawls JF
影响因子：
56.9
作者：
Shan, L;Molberg, O;Khosla, C
通讯作者：
Khosla, C
DOI：
10.1177/10454411930040030101
发表时间：
1993-01-01
期刊：
CRITICAL REVIEWS IN ORAL BIOLOGY & MEDICINE
影响因子：
--
作者：
LAMKIN, MS;OPPENHEIM, FG
通讯作者：
OPPENHEIM, FG
影响因子：
2.9
作者：
ABE, T;CHUNG, SI;FOLK, JE
通讯作者：
FOLK, JE