Laboratory of Food Technology and Leuven Food Science and Nutrition Research Centre (LFoRCe), Department of Microbial and Molecular Systems (M2S), Katholieke Universiteit Leuven, Kasteelpark Arenberg 22, Postbox 2457, 3001 Leuven, Belgium.
Planta. 2011 Feb;233(2):287-98. doi: 10.1007/s00425-010-1307-6. Epub 2010 Nov 3.
In order to gain insight into the in situ properties and localisation of kiwi pectin methylesterase inhibitor (PMEI), a toolbox of monoclonal antibodies (MA) towards PMEI was developed. Out of a panel of MA generated towards kiwi PMEI, three MA, i.e. MA-KI9A8, MA-KI15C12 and MA-KI15G7, were selected. Thorough characterisation proved that these MA bind specifically to kiwi PMEI and kiwi PMEI in complex with plant PME and recognise a linear epitope on PMEI. Extract screening of green kiwi (Actinidia deliciosa) and gold kiwi (Actinidia chinensis) confirmed the potential use of these MA as probes to screen for PMEI in other sources. Tissue printing revealed the overall presence of PMEI in pericarp and columella of ripe kiwi fruit. Further analysis on the cellular level showed PMEI label concentrated in the middle lamella and in the cell-wall region near the plasmalemma. Intercellular spaces, however, were either completely filled or lined with label. In conclusion, the developed toolbox of antibodies towards PMEI can be used as probes to localise PMEI on different levels, which can be of relevance for plant physiologists as well as food technologists.
为了深入了解猕猴桃果胶甲酯酶抑制剂(PMEI)的原位性质和定位,开发了一套针对 PMEI 的单克隆抗体(MA)工具。在针对猕猴桃 PMEI 产生的 MA 中,选择了三种 MA,即 MA-KI9A8、MA-KI15C12 和 MA-KI15G7。深入的特性分析证明,这些 MA 特异性结合猕猴桃 PMEI 和与植物 PME 结合的猕猴桃 PMEI,并识别 PMEI 上的线性表位。对绿猕猴桃(Actinidia deliciosa)和金猕猴桃(Actinidia chinensis)的提取物筛选证实了这些 MA 作为探针在其他来源中筛选 PMEI 的潜在用途。组织印迹显示 PMEI 在成熟猕猴桃果皮和果心的整体存在。进一步在细胞水平上的分析表明,PMEI 标记集中在中层和靠近质膜的细胞壁区域。然而,细胞间隙要么完全充满标记,要么被标记物排列。总之,针对 PMEI 开发的抗体工具包可用作探针,在不同水平上定位 PMEI,这对植物生理学家和食品技术人员都具有重要意义。