淀粉样蛋白增强因子(AEF)的生化性质及细胞来源

K Alizadeh-Khiavi, Z Ali-Khan
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摘要

采用低离子强度酸性缓冲液Sephadex G-200和Benzamidine-Sepharose (BZ)凝胶层析对肺泡包虫囊肿(AHC)诱导的淀粉样蛋白增强因子(AEF)进行了部分纯化。bz -凝胶结合的AEF (AEF- bz)在小鼠生物实验中显示出AEF活性,SDS-PAGE显示对Hide powder blue的蛋白水解活性为2个主要肽段和3个次要肽段。用10 mM苯基甲基磺酰氟或20 mM对氯苯苯甲酸预处理AEF-BZ,使其体内生物活性和体外蛋白水解活性完全丧失。产生抗AEF多克隆抗体(AAA),该抗体经被动转移到小鼠体内,完全消除酪蛋白诱导或ahc诱导的AEF的生物活性。与正常小鼠血清结合的Sepharose凝胶吸收的AAA在ahc感染小鼠和老年退休小鼠的AE和aef阳性血清中形成一条共同的沉淀带,免疫染色显示,它与ahc感染小鼠的大多数脾和腹膜白细胞的细胞质颗粒组分结合。相反,只有少数正常小鼠白细胞呈阳性染色。我们认为AEF很可能是一种源自白细胞的丝氨酸/硫醇蛋白酶,其在淀粉样变期间细胞内和体液浓度显著增加。本文讨论了溶酶体蛋白酶和首次成功制备的抗AEF抗体的作用,以及AEF的起源及其在淀粉样蛋白形成中的生物学功能。
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Biochemical nature and cellular origin of amyloid enhancing factor (AEF) as determined by anti-AEF antibody.

Low ionic strength acidic buffer, Sephadex G-200 and Benzamidine-Sepharose (BZ) gel chromatography, have been used for the partial purification of alveolar hydatid cyst (AHC) induced amyloid enhancing factor (AEF). BZ-gel bound AEF (AEF-BZ) demonstrated AEF activity in the mouse bioassay, proteolytic activity against Hide powder azure showed two major and three minor peptides on SDS-PAGE. Pretreatment of AEF-BZ with 10 mM phenylmethylsulphonyl fluoride or 20 mM p-chloromercuribenzoic acid completely abolished its bioactivity in vivo and proteolytic activity in vitro. Polyclonal anti-AEF antibody (AAA) was generated which on passive transfer into mice completely abolished the bioactivity of both casein-induced, or AHC-induced AEF. The AAA absorbed on Sepharose gel conjugated to normal mouse serum developed one common precipitin band between AE and AEF-positive sera from AHC-infected and old retired mice and in immunostaining it bound to the cytoplasmic granular components of a majority of splenic and peritoneal leucocytes from AHC-infected mice. In contrast, only a few normal mouse leucocytes showed positive staining. We suggest that AEF, in all probability, is a serine/thiol protease of leucocyte origin whose intracellular and humoral concentrations increase significantly during amyloidosis. The role of lysosomal proteases and anti-AEF antibody which has been successfully generated for the first time is discussed with reference to the origin of AEF and its presumed biological function in amyloidogenesis.

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