Heparanases Produce Distinct Populations of Heparan Sulfate Glycosaminoglycans in Chinese Hamster Ovary Cells*

Heparanases Produce Distinct Populations of Heparan Sulfate Glycosaminoglycans in Chinese Hamster Ovary Cells*
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乙酰肝素酶在中国仓鼠卵巢细胞中产生不同的硫酸乙酰肝素糖胺聚糖群体*

DOI:
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发表时间:
1997
影响因子:
4.8
通讯作者:
K. Robson
K. Robson
中科院分区:
生物学2区
文献类型:
--
作者:
K. Bame;K. Robson

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一旦内化,细胞相关硫酸乙酰肝素蛋白聚糖通过内切糖基化酶或乙酰肝素酶的作用降解为短的糖胺聚糖。我们已经开始解决的问题,有多少乙酰肝素酶是负责这个过程中,通过分析短硫酸乙酰肝素链在体内产生的中国仓鼠卵巢(CHO)细胞乙酰肝素酶。从CHO细胞中纯化硫酸乙酰肝素短链,并在还原端用[3 H] NaBH 4标记。通过对乙酰肝素酶链的单糖水解和对3 H-糖醇的分析表明,CHO细胞中的乙酰肝素酶活性为内切-β-葡萄糖醛酸苷酶。通过用亚硝酸或细菌肝素裂解酶处理[3 H]硫酸乙酰肝素链(其在特定序列处切割该链)并通过P2凝胶过滤色谱分析产物来检查乙酰肝素酶衍生的糖胺聚糖的修饰状态。两个群体的短链被确定为不同的程度上的乙酰肝素酶切割位点的非还原侧的修改。一类链从还原末端起至少9个残基是未修饰的,而另一类链在从乙酰肝素酶切割位点起3-7个残基内具有修饰的结构域。我们的研究结果表明,乙酰肝素酶的作用模型,其中的酶识别的硫酸盐含量的差异之间的修饰和未修饰的区域和结合的网站,包括这两个域。然后酶在修饰和未修饰序列之间的连接处切割糖胺聚糖,以产生不同的硫酸乙酰肝素短链群体。
Once internalized, cell-associated heparan sulfate proteoglycans are degraded to short glycosaminoglycans by the action of endoglycosidases or heparanases. We have begun to address the question of how many heparanases are responsible for this process by analyzing short heparan sulfate chains produced in vivo by Chinese hamster ovary (CHO) cell heparanases. Short heparan sulfate chains were purified from CHO cells and labeled at the reducing end with [3H]NaBH4. Hydrolysis of the chains to monosaccharides and analysis of the 3H-sugar alcohols indicate that heparanase activities in CHO cells are endo-β-glucuronidases. The modification state of the heparanase-derived glycosaminoglycans was examined by treating the [3H]heparan sulfate chains with nitrous acid or bacterial heparin lyases, which cut the chain at specific sequences, and analyzing the products by P2 gel filtration chromatography. Two populations of short chains were identified that differ in the extent of modification on the nonreducing side of the heparanase cleavage site. One class of chains is unmodified for at least 9 residues from the reducing end, while the other group has a modified domain within 3-7 residues from the heparanase cleavage site. Our results suggest a model of heparanase action where the enzymes recognize differences in sulfate content between modified and unmodified regions and bind to sites that encompass both domains. The enzymes then cleave the glycosaminoglycan at junctions between the modified and unmodified sequences to produce the different populations of short heparan sulfate chains.
DOI: 10.1016/s0021-9258(17)43350-3
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影响因子: --
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硫酸乙酰肝素 N-磺基转移酶缺陷的中国仓鼠卵巢细胞突变体中硫酸不足的硫酸乙酰肝素。
DOI: --
发表时间: 1989
期刊: The Journal of biological chemistry
影响因子: --
作者:
Bame,KJ;Esko,JD
通讯作者: Esko,JD
碱性成纤维细胞生长因子通过受体介导和硫酸乙酰肝素介导的机制内化。
DOI: --
发表时间: 1992
期刊: The Journal of biological chemistry
影响因子: --
作者:
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中国仓鼠卵巢细胞中的蛋白聚糖释放硫酸乙酰肝素糖胺聚糖不需要核心蛋白的蛋白水解。
DOI: --
发表时间: 1993
期刊: The Journal of biological chemistry
影响因子: --
作者:
Bame,KJ
通讯作者: Bame,KJ