ENDOTOXIN BINDING PROTEIN OF LIMULUS AMEBOCYTE

鲎变形细胞的内毒素结合蛋白

• 批准号: 3811040
• 负责人: C MINETTI
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3811040
• 关键词: affinity chromatography; bactericidal immunity; binding proteins; biological products; endotoxins; gel; horseshoe crabs; ion exchange chromatography; protein purification; pyrogens

Limulus amebocyte lysate forms a gel when exposed to minute quantities of endotoxin. This.unique property allowed the development of a sensitive assay for the detection of contaminating pyrogens in biological products. Our laboratory has been investigating the enzymatic mechanism of the lysate gelation. In the current study, using affinity columns of endotoxins and ion exchange chromatography a 12 kD protein with high affinity to endotoxins was isolated and purified. Binding studies with I-125-labeled 12 kD protein showed that the binding required the integrity of the endotoxin molecule to form the complex and that the lipid A moiety is essential but not sufficient to form the complex by itself. The gene coding for the 12 kD protein has also been isolated and sequenced. It contains a 31 amino acids signal sequence and a coding region for additional 104 amino acids. We propose that the 12 kD protein is one of the endotoxin binding proteins which form the defense mechanism to protect HSC against bacterial infections.

鲎变形细胞溶解物在暴露于微量的 内毒素这种独特的性质使敏感的 用于检测生物制品中污染热原的测定法。 我们实验室一直在研究裂解物的酶促机制 凝胶化在目前的研究中，使用内毒素和 离子交换色谱法，12 kD蛋白具有高亲和力， 分离并纯化内毒素。用I-125标记的12进行的结合研究 kD蛋白表明结合需要内毒素的完整性 分子形成复合物，脂质A部分是必需的， 不足以形成复合体本身。编码12 kD的基因 蛋白质也被分离和测序。它含有31种氨基酸 信号序列和另外104个氨基酸的编码区。我们 推测12 kD蛋白是内毒素结合蛋白之一 形成保护HSC免受细菌侵袭的防御机制 感染.