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通过硼酸亲和色谱法分离的膜相关硼相互作用蛋白。

Membrane-associated, boron-interacting proteins isolated by boronate affinity chromatography.

作者信息

Wimmer Monika A, Lochnit Günter, Bassil Elias, Mühling Karl H, Goldbach Heiner E

机构信息

Institute of Crop Science and Resource Conservation, University of Bonn, Bonn, Germany.

出版信息

Plant Cell Physiol. 2009 Jul;50(7):1292-304. doi: 10.1093/pcp/pcp073. Epub 2009 May 28.

Abstract

Boron deficiency symptoms point to a role for boron in plant membranes, but the molecular partners interacting with boron have not yet been identified. The objective of the present study was to isolate and identify membrane-associated proteins with an ability to interact with boron. Boron-interacting proteins were isolated from root microsomal preparations of arabidopsis (Arabidopsis thaliana) and maize (Zea mays) using phenylboronate affinity chromatography, subsequently separated by two-dimensional gel electrophoresis and identified using MALDI-TOF (matrix-assisted laser desorption ionization-time of flight) peptide mass fingerprinting. Twenty-six boron-binding membrane-associated proteins were identified in A. thaliana, and nine in Z. mays roots. Additional unidentified proteins were also present. Common to both species were the beta-subunit of mitochondrial ATP synthase, several beta-glucosidases, a luminal-binding protein and fructose bisphosphate aldolase. In A. thaliana, binding of these proteins to boron was significantly reduced after 4 d of boron deprivation. The relatively high number of diverse proteins identified as boron interacting, many of which are usually enriched in membrane microdomains, supports the hypothesis that boron plays a role in plant membranes by cross-linking glycoproteins, and may be involved in their recruitment to membrane microdomains.

摘要

硼缺乏症状表明硼在植物膜中发挥作用,但尚未鉴定出与硼相互作用的分子伴侣。本研究的目的是分离和鉴定具有与硼相互作用能力的膜相关蛋白。使用苯基硼酸亲和色谱法从拟南芥(Arabidopsis thaliana)和玉米(Zea mays)的根微粒体制备物中分离出硼相互作用蛋白,随后通过二维凝胶电泳进行分离,并使用基质辅助激光解吸电离飞行时间(MALDI-TOF)肽质量指纹图谱进行鉴定。在拟南芥中鉴定出26种与硼结合的膜相关蛋白,在玉米根中鉴定出9种。还存在其他未鉴定的蛋白质。两个物种共有的是线粒体ATP合酶的β亚基、几种β-葡萄糖苷酶、一种腔内结合蛋白和果糖二磷酸醛缩酶。在拟南芥中,缺硼4天后这些蛋白与硼的结合显著减少。被鉴定为与硼相互作用的多种蛋白质数量相对较多,其中许多通常富集在膜微区,这支持了硼通过交联糖蛋白在植物膜中发挥作用的假说,并且可能参与它们向膜微区的募集。

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