Peptidylarginine deiminase from Porphyromonas gingivalis citrullinates human fibrinogen and α-enolase: implications for autoimmunity in rheumatoid arthritis.
Peptidylarginine deiminase from Porphyromonas gingivalis citrullinates human fibrinogen and α-enolase: implications for autoimmunity in rheumatoid arthritis.
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DOI：
10.1002/art.27552
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发表时间：
2010-09
影响因子：
--
通讯作者：
Venables, Patrick J.
中科院分区：
文献类型：
--
作者：
Wegner, Natalia;Wait, Robin;Sroka, Aneta;Eick, Sigrun;Nguyen, Ky-Anh;Lundberg, Karin;Kinloch, Andrew;Culshaw, Shauna;Potempa, Jan;Venables, Patrick J.
研究牙周病原体牙龈卟啉单胞菌（牙龈卟啉单胞菌）的瓜氨酸蛋白作为破坏类风湿关节炎（RA）对瓜氨酸化蛋白耐受性的潜在机制。通过免疫印迹法分析牙龈卟啉单胞菌和其他10种口腔细菌的细胞提取物的内源性瓜氨酸化蛋白的表达。创建缺乏细菌PAD或牙龈卟啉单胞菌蛋白酶的牙龈卟啉单胞菌敲除菌株以评估这些酶在瓜氨酸中的作用。通过将活的野生型和敲除蛋白与牙龈卟啉单胞菌的人纤维蛋白原和α-烯醇酶一起孵育，并通过免疫印迹和质谱分析产物，研究了牙龈卟啉单胞菌对瓜氨酸酶的作用。内源性瓜氨酸蛋白在牙龈卟啉单胞菌中含量丰富，但在其他口腔细菌中缺乏。细菌PAD基因的缺失导致瓜氨酸蛋白质的完全废除。精氨酸-牙龈卟啉菌蛋白酶的失活，而不是赖氨酸-牙龈卟啉菌蛋白酶的失活，导致瓜氨酸减少。将野生型牙龈卟啉单胞菌与纤维蛋白原或α-烯醇化酶孵育，导致所得肽的羧基末端精氨酸残基处的蛋白质和瓜氨酸降解，通过质谱法鉴定。我们证明，牙龈卟啉单胞菌在测试的口腔细菌病原体中具有独特的瓜氨酸化蛋白的能力。我们进一步表明，牙龈卟啉单胞菌快速产生瓜氨酸化的宿主肽，其通过在精氨酸-X肽键处由精氨酸-牙龈卟啉酶进行蛋白水解裂解，然后通过细菌PAD对羧基末端精氨酸进行瓜氨酸化。我们的研究结果提示了一种新的模型，其中牙龈卟啉单胞菌介导的细菌和宿主蛋白的瓜氨酸酶提供了产生驱动RA中自身免疫反应的抗原的分子机制。
To investigate protein citrullination by the periodontal pathogen Porphyromonas gingivalis (P. gingivalis) as a potential mechanism for breaking tolerance to citrullinated proteins in rheumatoid arthritis (RA). Expression of endogenous citrullinated proteins was analysed by immunoblotting of cell extracts from P. gingivalis and ten other oral bacteria. P. gingivalis knockout strains lacking the bacterial PAD or gingipains were created to assess the role of these enzymes in citrullination. Citrullination of human fibrinogen and α-enolase by P. gingivalis was studied by incubating live wild-type and knockouts with the proteins and analysing the products by immunoblotting and mass spectrometry. Endogenous protein citrullination was abundant in P. gingivalis but lacking in the other oral bacteria. Deletion of the bacterial PAD gene resulted in complete abrogation of protein citrullination. Inactivation of arginine-gingipains, but not lysine-gingipains, led to decreased citrullination. Incubation of wild-type P. gingivalis with fibrinogen or α-enolase caused degradation of the proteins and citrullination of the resulting peptides at carboxy-terminal arginine residues, which were identified by mass spectrometry. We demonstrate that P. gingivalis is unique amongst the tested oral bacterial pathogens in its ability to citrullinate proteins. We further show that P. gingivalis rapidly generates citrullinated host peptides by proteolytic cleavage at arginine-X peptide bonds by arginine-gingipains followed by citrullination of carboxy-terminal arginines by bacterial PAD. Our results suggest a novel model where P. gingivalis-mediated citrullination of bacterial and host proteins provides a molecular mechanism for generating antigens driving the autoimmune response in RA.
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影响因子：
3.1
作者：
Jagels, MA;Travis, J;Hugli, TE
通讯作者：
Hugli, TE
影响因子：
4.9
作者：
Kinloch, Andrew;Tatzer, Verena;Wait, Robin;Peston, David;Lundberg, Karin;Donatien, Phillipe;Moyes, David;Taylor, Peter C;Venables, Patrick J
通讯作者：
Venables, Patrick J
影响因子：
3.1
作者：
IMAMURA, T;POTEMPA, J;TRAVIS, J
通讯作者：
TRAVIS, J
影响因子：
9.4
作者：
Griffen, AL;Lyons, SR;Leys, EJ
通讯作者：
Leys, EJ
影响因子：
4.4
作者：
Chapuy-Regaud, S;Sebbag, M;Serre, G
通讯作者：
Serre, G