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通过疏水亲和色谱法分离人补体因子H的两个分子群体。

Isolation of two molecular populations of human complement factor H by hydrophobic affinity chromatography.

作者信息

Ripoche J, Al Salihi A, Rousseaux J, Fontaine M

出版信息

Biochem J. 1984 Jul 1;221(1):89-96. doi: 10.1042/bj2210089.

DOI:10.1042/bj2210089
PMID:6235808
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1144006/
Abstract

Human complement factor H was prepared in highly purified form from fresh serum by euglobulin precipitation, DEAE-Sephacel chromatography and Sephacryl S-300 gel filtration. This preparation allowed the recovery of 37% of the initial factor H. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis revealed that factor H was homogeneous both in reduced and non-reduced media and exhibited a molecular mass of 150 kDa. Charge-shift experiments clearly showed the presence of hydrophobic sites in the factor H molecule. Charge shifts were observed with two detergent systems (Triton/sodium deoxycholate and Triton/cetyltrimethylammonium bromide). Factor H was able to bind to phenyl-Sepharose. This property allowed us to study two populations of factor H. These two populations exhibited the same physicochemical parameters, but revealed differences in their ability to aggregate in low- and iso-ionic-strength media. The molecular basis and biological significance of this heterogeneity are discussed.

摘要

通过优球蛋白沉淀、DEAE-葡聚糖凝胶层析和Sephacryl S-300凝胶过滤从新鲜血清中以高纯度形式制备人补体因子H。该制备方法可回收37%的初始因子H。十二烷基硫酸钠/聚丙烯酰胺凝胶电泳显示，因子H在还原和非还原介质中均为均一的，分子量为150 kDa。电荷转移实验清楚地表明因子H分子中存在疏水位点。在两种去污剂体系（Triton/脱氧胆酸钠和Triton/十六烷基三甲基溴化铵）中观察到了电荷转移。因子H能够结合到苯基琼脂糖上。这一特性使我们能够研究因子H的两个群体。这两个群体表现出相同的物理化学参数，但在低离子强度和等离子强度介质中聚集能力存在差异。讨论了这种异质性的分子基础和生物学意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7b3c/1144006/d7e30ff19712/biochemj00324-0097-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7b3c/1144006/9b8faf693611/biochemj00324-0096-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7b3c/1144006/924b64acd294/biochemj00324-0097-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7b3c/1144006/d7e30ff19712/biochemj00324-0097-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7b3c/1144006/9b8faf693611/biochemj00324-0096-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7b3c/1144006/924b64acd294/biochemj00324-0097-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7b3c/1144006/d7e30ff19712/biochemj00324-0097-b.jpg

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