Identification of multiple salicylic acid-binding proteins using two high throughput screens.
Identification of multiple salicylic acid-binding proteins using two high throughput screens.
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DOI：
10.3389/fpls.2014.00777
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发表时间：
2014
影响因子：
5.6
通讯作者：
Klessig DF
中科院分区：
文献类型：
--
作者：
Manohar M;Tian M;Moreau M;Park SW;Choi HW;Fei Z;Friso G;Asif M;Manosalva P;von Dahl CC;Shi K;Ma S;Dinesh-Kumar SP;O'Doherty I;Schroeder FC;van Wijk KJ;Klessig DF
水杨酸（SA）是一种重要的植物激素，参与植物成花诱导、气孔关闭、种子萌发、不定根发生和产热等过程。它还在响应非生物和生物胁迫过程中发挥关键作用。SA在信号传导抗病性中的作用是迄今为止研究得最好的过程，尽管它仍然只是部分理解。为了深入了解SA如何执行其各种功能，特别是在激活抗病性方面，开发了两种新的高通量筛选以鉴定新型SA结合蛋白（SABP）。第一个利用光反应性SA类似物4-AzidoSA（4AzSA）与拟南芥叶提取物中的蛋白质的交联，然后用抗SA抗体进行免疫选择，然后基于质谱的鉴定。第二个利用4AzSA与蛋白质微阵列（PMA）上的蛋白质的光亲和交联，随后用抗SA抗体检测。为了确定从这些筛选中获得的候选SABP（cSABP）是否是真正的SABP，产生重组产生的蛋白质，并测试与4AzSA的SA可识别交联，其通过免疫印迹分析监测，SA衍生物3-氨基乙基SA（3AESA）的SA可识别结合，其通过表面等离子体共振（SPR）测定检测，或[3 H]SA的SA可识别结合，其通过尺寸排阻色谱法检测。根据我们的标准，真正的SABP必须表现出SA结合活性，在至少两个这些测定，九个新的SABP确定在这里，其他九个以前报道。大约80个cSABP等待进一步评估。此外，关于NPR 1是否是SABP的矛盾报告通过显示其在上述所有三种测定中结合SA来解决。
Salicylic acid (SA) is an important hormone involved in many diverse plant processes, including floral induction, stomatal closure, seed germination, adventitious root initiation, and thermogenesis. It also plays critical functions during responses to abiotic and biotic stresses. The role(s) of SA in signaling disease resistance is by far the best studied process, although it is still only partially understood. To obtain insights into how SA carries out its varied functions, particularly in activating disease resistance, two new high throughput screens were developed to identify novel SA-binding proteins (SABPs). The first utilized crosslinking of the photo-reactive SA analog 4-AzidoSA (4AzSA) to proteins in an Arabidopsis leaf extract, followed by immuno-selection with anti-SA antibodies and then mass spectroscopy-based identification. The second utilized photo-affinity crosslinking of 4AzSA to proteins on a protein microarray (PMA) followed by detection with anti-SA antibodies. To determine whether the candidate SABPs (cSABPs) obtained from these screens were true SABPs, recombinantly-produced proteins were generated and tested for SA-inhibitable crosslinking to 4AzSA, which was monitored by immuno-blot analysis, SA-inhibitable binding of the SA derivative 3-aminoethylSA (3AESA), which was detected by a surface plasmon resonance (SPR) assay, or SA-inhibitable binding of [3H]SA, which was detected by size exclusion chromatography. Based on our criteria that true SABPs must exhibit SA-binding activity in at least two of these assays, nine new SABPs are identified here; nine others were previously reported. Approximately 80 cSABPs await further assessment. In addition, the conflicting reports on whether NPR1 is an SABP were addressed by showing that it bound SA in all three of the above assays.
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影响因子：
56.9
作者：
CHEN, ZX;SILVA, H;KLESSIG, DF
通讯作者：
KLESSIG, DF
DOI：
10.1073/pnas.88.18.8179
发表时间：
1991-09-01
影响因子：
11.1
作者：
CHEN, ZX;KLESSIG, DF
通讯作者：
KLESSIG, DF
影响因子：
56.9
作者：
METRAUX, JP;SIGNER, H;INVERARDI, B
通讯作者：
INVERARDI, B
影响因子：
11.6
作者：
CAO, H;BOWLING, SA;DONG, XN
通讯作者：
DONG, XN
DOI：
10.1073/pnas.92.14.6602
发表时间：
1995-07-03
影响因子：
11.1
作者：
DELANEY, TP;FRIEDRICH, L;RYALS, JA
通讯作者：
RYALS, JA