Differential profiles of salivary proteins with affinity to Streptococcus mutans lipoteichoic acid in caries-free and caries-positive human subjects
Differential profiles of salivary proteins with affinity to Streptococcus mutans lipoteichoic acid in caries-free and caries-positive human subjects
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DOI：
10.1111/omi.12057
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发表时间：
2014-10-01
影响因子：
3.7
通讯作者：
Han, S. H.
中科院分区：
文献类型：
--
作者：
Hong, S. W.;Seo, D. -G.;Han, S. H.
变形链球菌是引起龋齿和牙髓炎的代表性口腔病原体。它的脂磷壁酸（Sm.LTA）是一种重要的细胞壁毒力因子，参与细菌粘附和诱导炎症。由于Sm. LTA结合蛋白（Sm.LTA-BPs）可能在致病和宿主免疫中起重要作用，我们采用Sm. LTA结合微球和LTQ-Orbitrap混合傅立叶变换质谱法鉴定了无龋和有龋受试者唾液中的Sm.LTA-BPs。将Sm.LTA与N-羟基琥珀酰亚胺基-琼脂糖凝胶（（R））4快速流动珠（Sm. LTA-珠）缀合。Sm.LTA在结合过程中保留了其生物学特性，如通过在鼠巨噬细胞系中表达一氧化氮和干扰素诱导蛋白10以及在CHO/CD 14/TLR 2细胞中激活Toll样受体2（TLR 2）所确定的。Sm. LTA-BP从10名无龋或龋阳性的人类受试者的混合唾液中分离，进行测序以观察它们在各组中的差异表达，并通过高分辨率质谱法进一步鉴定。在无龋和龋阳性受试者的合并唾液中分别鉴定出8和12个具有统计学显著性的Sm. LTA-BP。无龋唾液中发现的独特Sm. LTA-BP包括组蛋白H4、profilin-1和中性粒细胞防御素-1，而在龋阳性唾液中发现的独特Sm. LTA-BP包括半胱氨酸蛋白酶抑制剂-C、半胱氨酸蛋白酶抑制剂-SN、半胱氨酸蛋白酶抑制剂-S、半胱氨酸蛋白酶抑制剂-D、溶菌酶C、钙调蛋白样蛋白3和-肌动蛋白。在两组中发现的Sm. LTA-BP是血红蛋白亚基和催乳素诱导蛋白、蛋白S100-A9和SPLUNC 2。总的来说，我们确定了Sm. LTA-BP在无龋和龋阳性受试者的唾液中，表现出差异蛋白质谱。
Streptococcus mutans is a representative oral pathogen that causes dental caries and pulpal inflammation. Its lipoteichoic acid (Sm.LTA) is known to be an important cell-wall virulence factor involved in bacterial adhesion and induction of inflammation. Since Sm.LTA-binding proteins (Sm.LTA-BPs) might play an important role in pathogenesis and host immunity, we identified the Sm.LTA-BPs in the saliva of caries-free and caries-positive human subjects using Sm.LTA-conjugated beads and LTQ-Orbitrap hybrid Fourier transform mass spectrometry. Sm.LTA was conjugated to N-hydroxysuccinimidyl-Sepharose((R)) 4 Fast Flow beads (Sm.LTA-beads). Sm.LTA retained its biological properties during conjugation, as determined by the expression of nitric oxide and interferon--inducible protein 10 in a murine macrophage cell line and activation of Toll-like receptor 2 (TLR2) in CHO/CD14/TLR2 cells. Sm.LTA-BPs were isolated from pooled saliva prepared from 10 caries-free or caries-positive human subjects each, electrophoresed to see their differential expression in each group, and further identified by high-resolution mass spectrometry. A total of 8 and 12 Sm.LTA-BPs were identified with statistical significance in the pooled saliva from the caries-free and caries-positive human subjects, respectively. Unique Sm.LTA-BPs found in caries-free saliva included histone H4, profilin-1 and neutrophil defensin-1, and those in caries-positive saliva included cystatin-C, cystatin-SN, cystatin-S, cystatin-D, lysozyme C, calmodulin-like protein 3 and -actin. The Sm.LTA-BPs found in both groups were hemoglobin subunits and , prolactin-inducible protein, protein S100-A9, and SPLUNC2. Collectively, we identified Sm.LTA-BPs in the saliva of caries-free and caries-positive subjects, which exhibit differential protein profiles.