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针对短鳍鳗鲡(澳洲鳗鲡)载脂蛋白B的抗血清的研制与部分特性鉴定

Development and partial characterisation of an antiserum against apolipoprotein B of the short-finned eel, Anguilla australis.

作者信息

Damsteegt Erin L, Mizuta Hiroko, Ozaki Yuichi, Hiramatsu Naoshi, Todo Takashi, Hara Akihiko, Ijiri Shigeho, Adachi Shinji, Lokman P Mark

机构信息

Department of Zoology, University of Otago, 340 Great King Street, PO Box 56, Dunedin, 9054, New Zealand,

出版信息

J Comp Physiol B. 2014 Jul;184(5):589-99. doi: 10.1007/s00360-014-0821-4. Epub 2014 Mar 11.

Abstract

Despite its key role in transportation of triacylglycerides in blood, the distribution, localisation and molecular weight variants of apolipoprotein B (Apob) in teleost fish have essentially escaped study. To address this, a specific short-finned eel (Anguilla australis) Apob antiserum was produced by an immunised rabbit, purified and partially characterised. Localisation of Apob at both the mRNA (in situ hybridisation) and protein (immunohistochemistry) levels mirrored that of mammals; thus immunostaining was confined to the interstitial spaces of the liver and the vascular core of the intestinal villi. Immunostaining of proteins by Western blotting, followed by high-resolution LC-MS, indicated that peptide sequence coverage of Apob in low-density lipoproteins spanned the full-length protein. We conclude that only full-length Apob is produced by eels and that both liver and intestine are key sites for its synthesis.

摘要

尽管载脂蛋白B(Apob)在硬骨鱼血液中三酰甘油的运输中起着关键作用,但其在硬骨鱼中的分布、定位和分子量变体基本上尚未得到研究。为了解决这个问题,通过免疫兔子制备了一种特异性的短鳍鳗(澳洲鳗鲡)Apob抗血清,进行了纯化并进行了部分特性鉴定。Apob在mRNA(原位杂交)和蛋白质(免疫组织化学)水平上的定位与哺乳动物相似;因此,免疫染色局限于肝脏的间质空间和肠绒毛的血管核心。通过蛋白质免疫印迹法进行蛋白质免疫染色,随后进行高分辨率液相色谱-质谱分析,结果表明低密度脂蛋白中Apob的肽序列覆盖了全长蛋白质。我们得出结论,鳗鱼只产生全长Apob,肝脏和肠道都是其合成的关键部位。

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