GLYCOPROTEIN-115, A GLYCOPROTEIN ISOLATED FROM CHICK BLOOD-VESSELS, IS WIDELY DISTRIBUTED IN CONNECTIVE-TISSUE
GLYCOPROTEIN-115, A GLYCOPROTEIN ISOLATED FROM CHICK BLOOD-VESSELS, IS WIDELY DISTRIBUTED IN CONNECTIVE-TISSUE
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DOI：
10.1083/jcb.100.1.18
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发表时间：
1985-01-01
影响因子：
7.8
通讯作者：
VOLPIN, D
中科院分区：
文献类型：
--
作者：
COLOMBATTI, A;BRESSAN, GM;VOLPIN, D
从鸡主动脉中分离出一种表观分子量为115,000的细胞外糖蛋白（gp 115）用于小鼠免疫。结果表明，该抗血清通过多种标准特异性识别gp 115：在主动脉提取物上，免疫印迹可以看到MW = 115,000左右的主要条带和较低MW的次要条带，并且在单克隆抗体上观察到类似的模式；与其他细胞外蛋白（纤连蛋白、层粘连蛋白、ⅰ型、ⅲ型、ⅳ型、ⅴ型和ⅵ型胶原）放射免疫结合无交叉反应性。采用亲和纯化抗体研究抗原在新生雏鸡冷冻组织切片上的分布。在血管中发现了很强的免疫反应性。在消化道中，荧光染色既局限于肌肉层水平，也局限于绒毛基质。在骨骼肌和心肌中，染色与大结缔组织束和每条肌纤维周围的基质有关。在肾脏、旁支气管平滑肌细胞丰富区、门静脉间隙和肝窦内观察到强烈的荧光。表皮-真皮交界处未检出抗原；真皮的免疫反应表现为弥漫性纤维状。免疫印迹分析证实，免疫荧光法在各脏器中检测到的抗原确实为gp 115；与主动脉提取物一样，在多个组织中检测到MW = 115,000左右的大条带。抗体反应物质也被纳入体外培养的胚胎平滑肌细胞产生的细胞外基质中，并被组织成细原纤维网。
An extracellular glycoprotein (gp 115) with an apparent MW = 115,000 isolated from chick aortas was used to immunize mice. The antisera were shown to specifically recognize gp 115 by numerous criteria: a major band around MW = 115,000 plus minor bands of lower MW were visible by immunoblotting on aorta extracts, and a similar pattern was observed with a monoclonal antibody; no cross-reactivity was detected by radioimmunobinding with other extracellular proteins, i.e., fibronectin, laminin and collagen types I, III, IV, V and VI. Antigen distribution on frozen tissue sections from newborn chicks was investigated by using affinity-purified antibody. Strong immunoreactivity was always found in blood vessels. In the digestive tract, the fluorescent staining was localized both at the level of muscular layers and in the stromal matrix of the villi. Within skeletal muscle and myocardium, staining was associated with large connective tissue bundles and the matrix around each muscle fiber. Intense fluorescence was observed in the kidney, in smooth muscle cells rich areas of parabronchi, and within the portal space and along liver sinusoids. The antigen was not detected at the epidermal-dermal junction; immunoreactivity in the dermis was present as a diffuse fibrillar pattern. That the antigen detected by immunofluorescence in the various organs was indeed gp 115 was demonstrated by immunoblotting analysis; as in aorta extracts, a major band around MW = 115,000 was detected in several tissues. Antibody-reacting material was also incorporated into the extracellular matrix produced by embryo smooth muscle cells grown in vitro and was organized as a meshwork of fine fibrils.