Characterization of an Acyl-CoA-binding protein from Arabidopsis thaliana
Characterization of an Acyl-CoA-binding protein from Arabidopsis thaliana
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DOI：
10.1006/abbi.1996.0282
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发表时间：
1996-07-01
影响因子：
3.9
通讯作者：
Ohlrogge, JB
中科院分区：
文献类型：
--
作者：
Engeseth, NJ;Pacovsky, RS;Ohlrogge, JB
从拟南芥中获得了一个cDNA克隆，其编码与牛酰基辅酶A结合蛋白（ACBP）具有高序列同一性（57%）的含有92个氨基酸残基的蛋白质。该克隆的编码序列在大肠杆菌中表达，并纯化基因产物（10.4kDa）。重组A.拟南芥ACBP（rAthACBP）显示结合酰基-CoA酯并保护酰基-CoA免于被微粒体酰基-水解酶降解。针对rAthACBP产生的抗体识别天然拟南芥ACBP，并且还与许多其他植物ACBP（包括油菜（Brassica napus）ACBP）交叉反应。使用蛋白质印迹法检测了该基因产物在拟南芥和芸苔属的各种组织中的表达模式和水平。A.拟南芥组织含有3 - 143 μ g AthACBP g（-1）FW，取决于组织（0.4 - 14 nmol g（-1）FW）。发展B。在种子成熟过程中，油菜种子的ACBP水平增加了12倍（20 - 250 pg ACBP g（-1）FW）;最高浓度出现在三酰甘油积累的峰值附近（26 nmol g（-1）FW）。(C)出版社：Academic Press，Inc.
A cDNA clone was obtained from Arabidopsis thaliana that encodes a protein containing 92 amino acid residues with high sequence identity (57%) to bovine acyl-CoA-binding protein (ACBP). The coding sequence of this clone was expressed in Escherichia coli and the gene product (10.4 kDa) was purified. The recombinant A. thaliana ACBP (rAthACBP) was shown to bind acyl-CoA esters and protect acyl-CoAs from degradation by microsomal acyl-hydrolases. Antibodies that were raised to rAthACBP recognized the native Arabidopsis ACBP and also cross-reacted with a number of other plant ACBPs, including rapeseed (Brassica napus) ACBP. The pattern of expression and level of the gene product were examined in various tissues of Arabidopsis and Brassica using Western blotting. A. thaliana tissues contained between 3 and 143 mu g AthACBP g(-1) FW depending on the tissue (0.4 to 14 nmol g(-1) FW). Developing B. napus seeds underwent a 12-fold increase in ACBP levels during seed maturation (20 to 250 pg ACBP g(-1) FW); the highest concentration occurring near the peak of triacylglycerol accumulation (26 nmol g(-1) FW). (C) 1996 Academic Press, Inc.