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人肺成纤维细胞的硫酸乙酰肝素蛋白聚糖。不同膜、基质和分泌形式的存在。

Heparan sulfate proteoglycans of human lung fibroblasts. Occurrence of distinct membrane, matrix and secreted forms.

作者信息

Lories V, David G, Cassiman J J, Van den Berghe H

出版信息

Eur J Biochem. 1986 Jul 15;158(2):351-9. doi: 10.1111/j.1432-1033.1986.tb09758.x.

Abstract

Human lung fibroblasts (HLF) were labeled with 35SO2-4 for 48 h and extracted with a guanidinium chloride buffer. A fraction of the extracted heparan sulfate proteoglycan (HSPG) appeared micelle-associated. In the absence of detergent these HSPG eluted in the void volume of Sepharose CL2B columns. In the presence of detergent these HSPG were included in Sepharose CL2B (Kav = 0.55) and 4B (Kav = 0.3) columns. This type of HSPG was specifically associated with isolated HLF cell membranes, suggesting that it may represent a fraction of integral membrane proteoglycans. Most of the HSPG in the HLF monolayers, however, eluted in the included volume of Sepharose CL2B (Kav = 0.4) and CL4B (Kav = 0.1) columns in the absence of detergent. This type of HSPG was not affected by detergent and was specifically retained in 'extracellular matrix' preparations. The medium of HLF monolayers contained HSPG of similar Mr as the membrane-associated HSPG. Of these three distinct HSPG fractions only the membrane-associated form could be incorporated in liposomes, confirming that the HSPG in this fraction may be integral membrane components.

摘要

人肺成纤维细胞(HLF)用35SO2-4标记48小时,并用氯化胍缓冲液提取。提取的硫酸乙酰肝素蛋白聚糖(HSPG)的一部分呈现出与微团相关。在没有去污剂的情况下,这些HSPG在琼脂糖CL2B柱的空体积中洗脱。在有去污剂的情况下,这些HSPG被包含在琼脂糖CL2B(Kav = 0.55)和4B(Kav = 0.3)柱中。这种类型的HSPG与分离的HLF细胞膜特异性相关,表明它可能代表整合膜蛋白聚糖的一部分。然而,HLF单层中的大多数HSPG在没有去污剂的情况下在琼脂糖CL2B(Kav = 0.4)和CL4B(Kav = 0.1)柱的包含体积中洗脱。这种类型的HSPG不受去污剂影响,并特异性保留在“细胞外基质”制剂中。HLF单层的培养基中含有与膜相关HSPG分子量相似的HSPG。在这三种不同的HSPG组分中,只有膜相关形式可以掺入脂质体中,证实该组分中的HSPG可能是整合膜成分。

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