P159 is a proteolytically processed, surface adhesin of Mycoplasma hyopneumoniae:: defined domains of P159 bind heparin and promote adherence to eukaryote cells
P159 is a proteolytically processed, surface adhesin of Mycoplasma hyopneumoniae:: defined domains of P159 bind heparin and promote adherence to eukaryote cells
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DOI：
10.1111/j.1365-2958.2006.05139.x
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发表时间：
2006-05-01
影响因子：
3.6
通讯作者：
Djordjevic, SP
中科院分区：
文献类型：
--
作者：
Burnett, TA;Dinkla, K;Djordjevic, SP
猪肺炎支原体（Mycoplasma hyopeptidase，Mycoplasma hyopeptidase hy已知肝素可抑制M.猪呼吸道上皮纤毛。M.肝素细胞结合肝素，但肝素结合蛋白的特性是有限的。M.猪链球菌裂解物鉴定了27 kDa（P27）、110 kDa（P110）和52 kDa（P52）蛋白质，它们代表来自mhp 494的159 kDa（P159）蛋白质的不同区域。这些裂解片段位于表面，并存在于所有生长阶段。在纯化跨越P159的四种重组蛋白（F1（P159）、F2（P159）、F3（P159）和F4（P159））后，仅F3（P159）和F4（P159）以剂量依赖性方式结合肝素（Kd值分别为142.37 +/- 22.01 nM; 75.37 +/- 7.34 nM）。扫描电子显微镜研究表明M.肝素钠与猪肾上皮样细胞（PK 15细胞）紧密结合，但这些过程被过量的肝素和F4抑制（P159）。同样，用F2（P159）和F4（P159）包被的乳胶珠粘附并进入PK 15细胞，但肝素、F2（P159）和F4（P159）是抑制性的。这些结果表明，P159是一个后切割的，糖胺聚糖结合粘附M。下丘脑
Mycoplasma hyopneumoniae, the causative agent of porcine enzootic pneumonia, colonizes the respiratory cilia of affected swine causing significant economic losses to swine production worldwide. Heparin is known to inhibit adherence of M. hyopneumoniae to porcine respiratory epithelial cilia. M. hyopneumoniae cells bind heparin but the identity of the heparin-binding proteins is limited. Proteomic analysis of M. hyopneumoniae lysates identified 27 kDa (P27), 110 kDa (P110) and 52 kDa (P52) proteins representing different regions of a 159 kDa (P159) protein derived from mhp494. These cleavage fragments were surface located and present at all growth stages. Following purification of four recombinant proteins spanning P159 (F1(P159), F2(P159), F3(P159) and F4(P159)), only F3(P159) and F4(P159) bound heparin in a dose-dependent manner (K-d values 142.37 +/- 22.01 nM; 75.37 +/- 7.34 nM respectively). Scanning electron microscopic studies showed M. hyopneumoniae bound intimately to porcine kidney epithelial-like cells (PK15 cells) but these processes were inhibited by excess heparin and F4(P159). Similarly, latex beads coated with F2(P159) and F4(P159) adhered to and entered PK15 cells, but heparin, F2(P159) and F4(P159) was inhibitory. These findings indicate that P159 is a post-translationally cleaved, glycosaminoglycan-binding adhesin of M. hyopneumoniae.