Moller Isabel, Sørensen Iben, Bernal Adriana J, Blaukopf Claudia, Lee Kieran, Øbro Jens, Pettolino Filomena, Roberts Alison, Mikkelsen Jørn Dalgaard, Knox J Paul, Bacic Antony, Willats William G T
Institute of Molecular Biology, University of Copenhagen, DK-1353, Denmark.
Plant J. 2007 Jun;50(6):1118-28. doi: 10.1111/j.1365-313X.2007.03114.x.
We describe here a methodology that enables the occurrence of cell-wall glycans to be systematically mapped throughout plants in a semi-quantitative high-throughput fashion. The technique (comprehensive microarray polymer profiling, or CoMPP) integrates the sequential extraction of glycans from multiple organs or tissues with the generation of microarrays, which are probed with monoclonal antibodies (mAbs) or carbohydrate-binding modules (CBMs) with specificities for cell-wall components. The profiles generated provide a global snapshot of cell-wall composition, and also allow comparative analysis of mutant and wild-type plants, as demonstrated here for the Arabidopsis thaliana mutants fra8, mur1 and mur3. CoMPP was also applied to Physcomitrella patens cell walls and was validated by carbohydrate linkage analysis. These data provide new insights into the structure and functions of plant cell walls, and demonstrate the potential of CoMPP as a component of systems-based approaches to cell-wall biology.
我们在此描述一种方法,该方法能够以半定量高通量的方式系统地绘制整个植物中细胞壁聚糖的分布图。该技术(综合微阵列聚合物分析,即CoMPP)将从多个器官或组织中顺序提取聚糖与微阵列的生成相结合,然后用对细胞壁成分具有特异性的单克隆抗体(mAb)或碳水化合物结合模块(CBM)对微阵列进行探测。所生成的图谱提供了细胞壁组成的全局快照,还允许对突变体和野生型植物进行比较分析,如此处对拟南芥突变体fra8、mur1和mur3所展示的那样。CoMPP也应用于小立碗藓的细胞壁,并通过碳水化合物连接分析进行了验证。这些数据为植物细胞壁的结构和功能提供了新的见解,并证明了CoMPP作为基于系统的细胞壁生物学方法组成部分的潜力。