Isolation of apoproteins from canine surface active material.
Isolation of apoproteins from canine surface active material.
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从犬表面活性物质中分离脱辅基蛋白。
DOI：
10.1152/ajplegacy.1973.224.4.788
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发表时间：
1973
期刊：
影响因子：
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通讯作者：
J. Clements
中科院分区：
文献类型：
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作者：
R. J. King;D. Klass;E. Gikas;J. Clements
理查德·j·金、丹尼尔·j·克拉斯、埃利亚斯·g·吉卡斯和约翰·a·克莱门茨。犬表面活性物质载脂蛋白的分离。点。[j] .中国生物医学工程学报，2009(4):387 - 398。1973.我们从犬表面活性物质中分离出蛋白质，并将其溶解于水溶液中，研究其理化性质。其中三种蛋白经电泳和免疫学鉴定为白蛋白、IgG和非血清蛋白。最后一种蛋白的分子量为10- 11000，对磷脂有很强的亲和力，并且含有非常大比例的疏水氨基酸残基。定量免疫测定表明，该蛋白在纯化的表面活性物质中的浓度是整个肺匀浆的50倍。该蛋白的理化性质、免疫荧光在肺泡界面的定位以及在纯化表面活性物质中的浓度与其提出的肺表面活性物质特异性一致。肺表面活性物质;脂蛋白;脱辅基蛋白;dipalmitoyl磷脂酰胆碱;肺表面活性物质的化学组成已在几个实验室进行了研究（12,13,17,23 - 25,29,31），普遍认为它富含磷脂，特别是双棕榈酰磷脂酰胆碱（DPC）。与它相关的蛋白质是否有资格作为脂蛋白一直存在争议(9)。为了研究这个问题，我们从成年狗的气管灌洗液和全肺匀浆中分离出表面活性物质（16），并在37℃下测试了它们的表面特性。我们发现这些材料具有预测的肺泡原位表面活性剂的表面特性（17），并且它们总是含有超过9%的蛋白质。更详细地研究了从气管灌洗液中纯化的表面活性物质，我们发现它作为脂蛋白通过八次以上的离心、两次连续密度梯度、pH梯度等电聚焦、淀粉凝胶电泳和另外六次离心浮选（17）进行迁移。由于蛋白质和脂质成分的这种一致的关联以及蛋白质部分可能的生理重要性，我们研究了它们在犬表面活性物质中的身份和性质。
KING, RICHARD J., DANIEL J. KLASS, ELIAS G. GIKAS, AND JOHN A. CLEMENTS. Isolation of apoproteins from canine surface active material. Am. J. Physiol. 224 (4): 788-795. 1973.~ We isolated the proteins in canine surface active material, solubilized them in aqueous solutions, and studied their physical and chemical properties. Three of these proteins were identified electrophoretically and immunologically as albumin, IgG, and a non-serum protein. The last protein has a molecular weight of 10-l 1,000, a strong affinity for phospholipid, and a very large percentage of hydrophobic amino acid residues. As measured by quantitative immunoassay, this protein is concentrated 50-fold in purified surface active material, as compared with the whole homogenate of lung. The physical and chemical properties of this protein, its localization at the alveolar interface by immunofluorescence, and its concentration in purified surface active material are consistent with its proposed specificity for pulmonary surfactant. pulmonary surfactant; lipoprotein; apoprotein; dipalmitoyl phosphatidylcholine; immunoassayTHE CHEMICAL COMPOSITION Of pulmonary SUrfaCe active material has been investigated in several laboratories(12, 13, 17, 23-25, 29, 3 1), and there is general agreement that it is rich in phospholipids, especially dipalmitoyl phosphatidylcholine(DPC). Whether or not the protein associated with it qualifies it as a lipoprotein has been debated (9). In order to examine this question, we have isolated surface active materials from both tracheal lavage fluid and whole-lung homogenate of adult dogs (16) and have tested their surface properties at 37 C. We find that these materials have the surface properties predicted for the surfactant lining the alveoli in situ (17) and that invariably they contain over 9% protein. Studying in greater detail the surface active material purified from the tracheal lavage fluid, we find that it migrates as a lipoprotein through more than eight centrifugations, two continuous density gradients, isoelectric focusing in pH gradients, in starch gel electrophoresis, and during six additional centrifugal flotations (17). Because of this consistent association of the protein and lipid components and because of the possible physiological importance of the protein moieties, we have investigated their identity and properties in canine surface active material.