Institute of Dental Research, Westmead Millennium Institute and Centre for Oral Health, Westmead Hospital, Sydney, New South Wales, Australia.
Infect Immun. 2012 Oct;80(10):3733-41. doi: 10.1128/IAI.00528-12. Epub 2012 Jul 16.
In the context of periodontal disease, cysteine proteinases or gingipains from Porphyromonas gingivalis have been implicated in the hydrolysis of cytokines, including gamma interferon (IFN-γ). This cytokine plays a crucial role in host defenses, in part, by regulating expression of major histocompatibility complex molecules. Our recent analysis has identified three structurally defined modules, K1, K2, and K3, of the hemagglutinin region of the lysine gingipain Kgp. These three structurally homologous domains have a common β-sandwich topology that is similar to that found in a superfamily of adhesins and carbohydrate binding domains. The three Kgp hemagglutinin modules are distinguished by variation in some of the loops projecting from the β-sandwich core. Recombinant products corresponding to both single and multidomain regions as well as native Kgp bound IFN-γ with similar affinities. Among the adhesin domain constructs, only the K1K2 polypeptide inhibited the upregulation of HLA-1 expression in a human erythroleukemia (K562) line induced by both recombinant and native IFN-γ. The K1K2 polypeptide also inhibited HLA-DR expression induced by IFN-γ in human umbilical vein endothelial cells. These effects were competitively inhibited by coincubation with sodium or potassium chloride solution. The N-terminal residues of IFN-γ were implicated in mediating the effect of K1K2, while antibody binding to loop 1 of K2 blocked the action of K1K2. The findings indicate the potential significance of structurally defined Kgp adhesin modules in the inactivation of IFN-γ but also the potential of K1K2 in locating the target for the catalytic domain of Kgp.
在牙周病的背景下,牙龈卟啉单胞菌的半胱氨酸蛋白酶或牙龈蛋白酶被认为参与了细胞因子的水解,包括γ干扰素(IFN-γ)。这种细胞因子在宿主防御中起着至关重要的作用,部分原因是它调节主要组织相容性复合体分子的表达。我们最近的分析确定了赖氨酸牙龈蛋白酶 Kgp 的血凝素区域的三个结构定义的模块,K1、K2 和 K3。这三个结构同源的结构域具有共同的β-三明治拓扑结构,类似于一个超级家族的粘附素和碳水化合物结合结构域。这三个 Kgp 血凝素模块的区别在于从β-三明治核心伸出的一些环的变化。与单个和多结构域区域以及天然 Kgp 相对应的重组产物与 IFN-γ 具有相似的亲和力。在粘附素结构域构建体中,只有 K1K2 多肽抑制了重组和天然 IFN-γ诱导的人红白血病(K562)系 HLA-1 表达的上调。K1K2 多肽还抑制了 IFN-γ诱导的人脐静脉内皮细胞 HLA-DR 的表达。这些效应可通过与氯化钠或氯化钾溶液共孵育而被竞争性抑制。IFN-γ 的 N 末端残基参与介导 K1K2 的作用,而 K2 环 1 的抗体结合阻断了 K1K2 的作用。这些发现表明结构定义的 Kgp 粘附素模块在 IFN-γ失活中的潜在意义,但也表明 K1K2 在定位 Kgp 催化结构域的靶标方面的潜力。