Heparanase-mediated cleavage of macromolecular heparin accelerates release of granular components of mast cells from extracellular matrices
Heparanase-mediated cleavage of macromolecular heparin accelerates release of granular components of mast cells from extracellular matrices
复制标题
DOI：
10.1042/bj20131463
复制
发表时间：
2014-03-01
影响因子：
4.1
通讯作者：
Irimura, Tatsuro
中科院分区：
文献类型：
--
作者：
Higashi, Nobuaki;Waki, Michihiko;Irimura, Tatsuro
乙酰肝素酶裂解结缔组织型肥大细胞分泌颗粒中的大分子肝素。我们研究了在模拟肥大细胞生理学驻留的微环境下裂解的作用。结缔组织型肥大细胞系MST和小鼠腹膜细胞来源的肥大细胞在分泌颗粒中储存大分子肝素。由于肝素的乙酰肝素酶依赖性裂解，表达乙酰肝素酶的细胞储存了碎片化的肝素（类似于 10 kDa）。我们生产了一种基于人造胶原蛋白的细胞外基质，并将活细胞或从细胞中纯化的糖胺聚糖放入基质中，以测量硫酸化大分子向培养基中的释放。来自表达脱颗粒乙酰肝素酶的细胞和纯化的糖胺聚糖的硫酸盐放射性标记分子显示出比来自模拟细胞的那些分子显着更多的释放到培养基中，而悬浮培养中的情况并非如此。与模拟细胞相比，肥大细胞颗粒酶食糜酶（而非β-己糖胺酶）从脱颗粒乙酰肝素酶表达细胞中释放出显着更多的释放。与单独的酶或与来自模拟细胞的大分子肝素混合相比，纯化的食糜酶与来自乙酰肝素酶表达细胞的片段化肝素混合显示出更多的胶原蛋白凝胶释放。我们提出乙酰肝素酶对大分子肝素的裂解加速了肝素和食糜酶从细胞外基质的释放。
Heparanase cleaves macromolecular heparin in the secretory granules of connective tissue-type mast cells. We investigated roles of the cleavage under a microenvironment mimicking where the mast cells physiologically reside. A connective tissue-type mast cell line MST and mouse peritoneal cell-derived mast cells stored macromolecular heparin in the secretory granules. The cells expressing heparanase stored fragmented heparin (similar to 10 kDa) due to heparanase-dependent cleavage of the heparin. We produced an artificial collagen-based extracellular matrix and placed the live cells or glycosaminoglycans purified from the cells in the matrix to measure the release of sulfated macromolecules into the medium. The sulfate-radiolabelled molecules from the degranulating heparanase-expressing cells and the purified glycosaminoglycans showed significantly greater release into the medium than those derived from mock cells, which was not the case in suspension culture. The mast cell granular enzyme chymase, but not beta-hexosaminidase, showed significantly greater release from the degranulating heparanase-expressing cells than from mock cells. Purified chymase mixed with fragmented heparin derived from heparanase-expressing cells showed greater release from collagen gels than the enzyme alone or mixed with macromolecular heparin derived from mock cells. We propose that the cleavage of macromolecular heparin by heparanase accelerates the release of heparin and chymase from extracellular matrices.