Pacák F, Siegert R
Eur J Biochem. 1982 Oct;127(2):375-80. doi: 10.1111/j.1432-1033.1982.tb06882.x.
Crude pyrogen was obtained by stimulating 1.4 x 10(12) peritoneal-exudate leukocytes from rabbits in vitro with Escherichia coli endotoxin. A pyrogen molecule was isolated by ammonium-sulfate precipitation, polyacrylamide gel electrophoresis at pH 5.5 and 8.9, and gel filtration on Bio-Gel P-100. These procedures yielded a pyrogenic fraction which was electrophoretically and immunologically homogeneous. The pyrogenicity was 50 ng/kg of rabbit. This leukocyte pyrogen consisted of a protein with a molecular weight of approximately 14 000. Amino-acid analysis showed high amounts of glutamic acid and glycine; methionine and tryptophan were present in small quantities. Arginine was found to be the N-terminal amino acid. The molecule contained about 0.8% neutral sugar and 0.7% lipids. Its approximate isoelectric point in citrate/phosphate buffer was 4.2. The activity was lost by lyophilization and storage at - 18 degrees C for six months. The data presented here are the first detailed characterization of a rabbit leukocyte pyrogen. The properties in common to leukocytic pyrogen and interleukin 1 are discussed.
通过用大肠杆菌内毒素在体外刺激1.4×10¹²只兔腹腔渗出白细胞获得粗制热原。通过硫酸铵沉淀、pH 5.5和8.9的聚丙烯酰胺凝胶电泳以及在Bio-Gel P-100上进行凝胶过滤分离出热原分子。这些步骤产生了一个在电泳和免疫方面均一的致热部分。其致热活性为50 ng/kg兔。这种白细胞热原由一种分子量约为14000的蛋白质组成。氨基酸分析表明谷氨酸和甘氨酸含量高；蛋氨酸和色氨酸含量少。发现精氨酸是N端氨基酸。该分子含有约0.8%的中性糖和0.7%的脂质。其在柠檬酸盐/磷酸盐缓冲液中的近似等电点为4.2。冻干并在-18℃储存六个月后活性丧失。此处给出的数据是对兔白细胞热原的首次详细表征。讨论了白细胞热原和白细胞介素1的共同特性。
