Department of Ophthalmology, University of Helsinki, Finland.
J Lipid Res. 2010 Nov;51(11):3126-34. doi: 10.1194/jlr.M006239. Epub 2010 Aug 19.
In addition to circulation, where it transfers phospholipids between lipoprotein particles, phospholipid transfer protein (PLTP) was also identified as a component of normal tear fluid. The purpose of this study was to clarify the secretion route of tear fluid PLTP and elucidate possible interactions between PLTP and other tear fluid proteins. Human lacrimal gland samples were stained with monoclonal antibodies against PLTP. Heparin-Sepharose (H-S) affinity chromatography was used for specific PLTP binding, and coeluted proteins were identified with MALDI-TOF mass spectrometry or Western blot analysis. Immunoprecipitation assay and blotting with specific antibodies helped to identify and characterize PLTP-mucin interaction in tear fluid. Human tear fluid PLTP is secreted from the lacrimal gland. MALDI-TOF analysis of H-S fractions identified several candidate proteins, but protein-protein interaction assays revealed only ocular mucins as PLTP interaction partners. We suggest a dual role for PLTP in human tear fluid: (1) to scavenge lipophilic substances from ocular mucins and (2) to maintain the stability of the anterior tear lipid film. PLTP may also play a role in the development of ocular surface disease.
除了在脂蛋白颗粒之间转移磷脂外,磷脂转移蛋白 (PLTP) 也被鉴定为正常泪液的成分。本研究旨在阐明泪液 PLTP 的分泌途径,并阐明 PLTP 与其他泪液蛋白之间的可能相互作用。用人抗 PLTP 单克隆抗体对人泪腺组织进行染色。用肝素-琼脂糖 (H-S) 亲和层析法进行特异性 PLTP 结合,用 MALDI-TOF 质谱或 Western blot 分析鉴定共洗脱蛋白。免疫沉淀试验和用特异性抗体进行的印迹分析有助于鉴定和表征泪液中的 PLTP-粘蛋白相互作用。人泪液 PLTP 从泪腺分泌。H-S 级分的 MALDI-TOF 分析鉴定了几种候选蛋白,但蛋白-蛋白相互作用试验仅显示眼粘蛋白是 PLTP 的相互作用伙伴。我们提出 PLTP 在人泪液中具有双重作用:(1) 从眼粘蛋白中清除亲脂性物质,(2) 维持前泪液脂质膜的稳定性。PLTP 也可能在眼表面疾病的发展中起作用。