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从玉米（Zea mays）根细胞中提取具有Ca2+依赖性ATP酶活性的膜结合蛋白及其在体外膜融合中的可能作用。

The extraction from maize (Zea mays) root cells of membrane-bound protein with Ca2+-dependent ATPase activity and its possible role in membrane fusion in vitro.

作者信息

Baydoun E A, Northcote D H

出版信息

Biochem J. 1981 Mar 1;193(3):781-92. doi: 10.1042/bj1930781.

DOI:10.1042/bj1930781
PMID:6458276
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1162667/
Abstract

Membrane fusion in vitro between Golgi apparatus- and plasma-membrane-rich fractions isolated from maize (Zea mays) roots was found to be dependent on Ca2+ and the membrane proteins. Trypsin treatment of mixed membrane fractions before the addition of Ca2+ inhibited their ability to fuse. It resulted also in a selective and progressive elimination of a characteristic intense polypeptide band (B1) on gel electrophoresis. This polypeptide was not removed by chymotrypsin or thermolysin. B1 is an integral membrane protein with an exposed portion to the outside. Sodium deoxycholate was used to solubilize the proteins of mixed membrane fractions. Extracted proteins analysed by non-SDS (sodium dodecyl sulphate) polyacrylamide-gel electrophoresis revealed the presence of four isolated bands. When re-electrophoresed in the presence of SDS, one of these bands exhibited the same mobility as polypeptide B1. Enzymic staining of non-SDS-polyacrylamide gels showed that this protein has Ca2+- and Mg2+-dependent ATPase activity. Its possible role in membrane fusion is discussed.

摘要

研究发现，从玉米（Zea mays）根部分离出的富含高尔基体和质膜的组分在体外的膜融合依赖于Ca2+和膜蛋白。在添加Ca2+之前，用胰蛋白酶处理混合膜组分可抑制其融合能力。这也导致在凝胶电泳上选择性地逐步消除一条特征性的强多肽带（B1）。这条多肽不会被胰凝乳蛋白酶或嗜热菌蛋白酶去除。B1是一种整合膜蛋白，有一部分暴露在外部。用脱氧胆酸钠溶解混合膜组分中的蛋白质。通过非SDS（十二烷基硫酸钠）聚丙烯酰胺凝胶电泳分析提取的蛋白质，发现有四条分离的条带。当在SDS存在下重新电泳时，其中一条带的迁移率与多肽B1相同。非SDS-聚丙烯酰胺凝胶的酶染色显示，这种蛋白质具有Ca2+和Mg2+依赖性ATP酶活性。文中讨论了其在膜融合中可能的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/f27e807097d0/biochemj00406-0133-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/e4ab9fd8bb5d/biochemj00406-0129-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/60ed6d907206/biochemj00406-0129-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/841b29727692/biochemj00406-0130-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/cb434ff5702e/biochemj00406-0131-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/36cd2ced0b23/biochemj00406-0132-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/26cb4ce14851/biochemj00406-0133-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/4ed21ee8ba14/biochemj00406-0133-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/f27e807097d0/biochemj00406-0133-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/e4ab9fd8bb5d/biochemj00406-0129-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/60ed6d907206/biochemj00406-0129-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/841b29727692/biochemj00406-0130-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/cb434ff5702e/biochemj00406-0131-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/36cd2ced0b23/biochemj00406-0132-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/26cb4ce14851/biochemj00406-0133-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/4ed21ee8ba14/biochemj00406-0133-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d62/1162667/f27e807097d0/biochemj00406-0133-c.jpg

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本文引用的文献

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Purification and Characterization of a Cation-stimulated Adenosine Triphosphatase from Corn Roots.从玉米根中纯化和鉴定一种阳离子刺激的三磷酸腺苷酶。
Plant Physiol. 1978 Aug;62(2):165-72. doi: 10.1104/pp.62.2.165.
2
The Incorporation of d-Glucosamine into Glycolipids and Glycoproteins of Membrane Preparations from Phaseolus aureus Hypocotyls.金盏花下胚轴膜制剂糖脂和糖蛋白中 d-葡萄糖胺的掺入。
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A spectrophotometric determination of trypsin and chymotrypsin.胰蛋白酶和胰凝乳蛋白酶的分光光度测定法
Biochim Biophys Acta. 1955 Apr;16(4):570-5. doi: 10.1016/0006-3002(55)90280-8.
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Histochemistry of hepatic phosphatases of a physiologic pH; with special reference to the demonstration of bile canaliculi.生理pH值下肝脏磷酸酶的组织化学；特别提及胆小管的显示
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Measurement and characteristics of fusion of isolated membrane fractions from maize root tips.玉米根尖分离膜组分融合的测量与特性
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