Preparation of subcellular fractions from granulation tissue by density gradient centrifugation.

P Lehtinen, E Kulonen
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引用次数: 6

Abstract

The 7000 g pellet of homogenized mature sponge-induced granulation tissue was fractionated by centrifugation in a stepwise sucrose gradient in order to study the synthesis and secretion of collagen and other components of this tissue on the subcellular level. As indicated by chemical and enzymatic assays, by electron microscopy and by incorporation experiments, the collagen-synthethesizing rough endoplasmic reticulum fraction was isolated free from the secretory vesicles (smooth endoplasmic membranes and Golgi elements) and fibrous extracellular matrix. Collagen differed from other proteins in the distribution among the subcellular fractions. In pulse-chase experiment the translocation of 14C-labelled collagen was demonstrated from the rough endoplasmic reticulum through the secretory vesicles to extracellular fibrillar collagen. This fractionation method will be used to study the modulation of collagen synthesis and secretion in the reparative tissue.

密度梯度离心法制备肉芽组织亚细胞组分。
取匀浆后的成熟海绵肉芽组织7000 g颗粒,经蔗糖梯度离心分离,在亚细胞水平上研究该组织中胶原等成分的合成和分泌。通过化学和酶分析、电子显微镜和结合实验表明,胶原合成粗内质网部分从分泌囊泡(光滑内质膜和高尔基体)和纤维细胞外基质中分离出来。胶原蛋白在亚细胞组分中的分布与其他蛋白不同。脉冲追踪实验表明,14c标记的胶原蛋白从粗内质网经分泌囊泡向细胞外纤维性胶原转移。这种分离方法将用于研究修复组织中胶原合成和分泌的调节。
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