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Lipopolysaccharide-binding proteins of Limulus amebocyte lysate.

机译：am变形细胞溶解物的脂多糖结合蛋白。

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Limulus amebocyte lysate, obtained from horseshoe crab (Limulus polyphemus) blood cells, contains a coagulation system which is activated by bacterial lipopolysaccharide (LPS). A chromatographic fraction of Limulus lysate, containing the endotoxin-sensitive factor(s) which initiates the coagulation cascade, was studied. We utilized a photoreactive, cleavable, radiolabeled derivative of Salmonella minnesota LPS, LPS-(p-azidosalicylamido)-1,3'-dithiopropionamide (LPS-ASD), to identify LPS-binding proteins. The lysate fraction was incubated with LPS-ASD, and LPS-binding proteins were identified by autoradiography of sodium dodecyl sulfate-polyacrylamide gels. An 82-kDa protein, a major protein component of this fraction from Limulus lysate, was identified as a LPS-binding protein in a majority of lysates. Incubation of whole Limulus lysate with antiserum to this protein resulted in enhanced sensitivity of the lysate to LPS, suggesting that this 82-kDa protein is a negative regulator of coagulation. A minor 50-kDa protein component of lysate also was identified as a LPS-binding protein and is a candidate for the LPS-sensitive coagulation protein in L. polyphemus.
机译：从horse（blood）血细胞中获得的am变形细胞裂解物含有一个凝固系统，该凝固系统被细菌脂多糖（LPS）激活。对Li裂解物的色谱级分进行了分析，其中含有内毒素敏感因子，可引发凝血级联反应。我们利用光反应性，可裂解，放射性标记的明尼苏达沙门氏菌LPS，LPS-（对叠氮基水杨酰胺基）-1,3'-二硫代丙酰胺（LPS-ASD）的衍生物来识别LPS结合蛋白。将裂解物级分与LPS-ASD孵育，并通过放射自显影十二烷基硫酸钠-聚丙烯酰胺凝胶来鉴定LPS结合蛋白。来自Li裂解物的该级分的主要蛋白质组分82-kDa蛋白被鉴定为大多数裂解物中的LPS结合蛋白。整个Li裂解物与该蛋白的抗血清一起孵育会导致裂解物对LPS的敏感性增强，表明该82 kDa蛋白是凝血的负调节剂。溶胞产物中50kDa的次要蛋白质成分也被确定为LPS结合蛋白，并且是多角乳杆菌中LPS敏感凝血蛋白的候选者。

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