PLUNC in human nasal lavage fluid:: multiple isoforms that bind to lipopolysaccharide
PLUNC in human nasal lavage fluid:: multiple isoforms that bind to lipopolysaccharide
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DOI：
10.1016/j.bbapap.2004.01.001
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发表时间：
2004-06-01
影响因子：
3.2
通讯作者：
Lindahl, M
中科院分区：
文献类型：
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作者：
Ghafouri, B;Kihlström, E;Lindahl, M
在这里，我们证明了人鼻灌洗液（NLF）中存在腭肺鼻上皮克隆（PLUNC）的多种异构体。通过二维凝胶电泳（2-DE）分离出8个异构体，并使用基质辅助激光解吸/电离飞行时间质谱法（MALDI-TOF MS）对胰蛋白酶和天冬氨酸酶裂解进行肽图谱绘制。利用纳米电喷雾质谱（MS/MS）对碰撞诱导解离（CID）片段谱进行分析，并进行氨基酸测序验证。在n -糖苷酶处理后，一个异构体显示出电泳迁移率的变化，表明至少有一个PLUNC异构体被糖基化了。我们还证明了NLF中的PLUNC在体外与脂多糖（LPS）结合；事实上，在NLF中存在的所有蛋白质中，只有PLUNC亚型被发现吸附在lps包被的表面上。这些结果表明，PLUNC在人NLF中以多种lps结合亚型表达。推测PLUNC可能在上呼吸道先天免疫应答中发挥作用。(C) 2004 Elsevier B.V.版权所有
Here, we demonstrate the presence of multiple isoforms of palate lung nasal epithelial clone (PLUNC) in human nasal lavage fluid (NLF). Eight isoforms were separated by two-dimensional gel electrophoresis (2-DE), and peptide mapping of the proteins was performed using MALDI-TOF MS (matrix assisted laser desorption/ionization time of flight mass spectrometry) of tryptic and asparginase cleavages. The identification was verified by amino acid sequencing after analysis of collision-induced dissociation (CID) fragmentation spectra with nanoelectrospray MS/MS. One isoform showed an electrophoretic mobility shift after N-glycosidase treatment, indicating that at least one of the PLUNC isoforms is glycosylated. We also demonstrate that PLUNC in NLF binds to lipopolysaccharide (LPS) in vitro; indeed, out of all proteins present in NLF only the PLUNC isoforms were found to adsorb to an LPS-coated surface. These results show that PLUNC is expressed as multiple LPS-binding isoforms in human NLF. The possibility that PLUNC may play a role in the innate immune response of the upper airways is inferred. (C) 2004 Elsevier B.V. All rights reserved.