Abstract：
Aquaporin-0 (AQP0) constitutes 50 % of the lens membrane proteome and plays important roles in lens fiber cell adhesion, water permeability, and lens transparency. Previous work has shown that specific proteins, such as calmodulin (CaM), interact with AQP0 to modulate its water permeability; however, these studies often used AQP0 peptides, rather than full-length protein, to probe these interactions. Furthermore, the specific regions of interaction of several known AQP0 interacting partners, i.e. αA and αB-crystallins, and phakinin (CP49) remain unknown. The purpose of this study was to use crosslinking mass spectrometry (XL-MS) to identify interacting proteins with full-length AQP0 in crude lens cortical membrane fractions and to determine the specific protein regions of interaction. Our results demonstrate, for the first time, that the AQP0 N-terminus can engage in protein interactions. Specific regions of interaction are elucidated for several AQP0 interacting partners including phakinin, α-crystallin, connexin-46, and connexin-50. In addition, two new interacting partners, vimentin and connexin-46, were identified.
摘要：
水通道蛋白-0(AQP0)占晶状体膜蛋白质组的50%,在晶状体成纤维细胞粘附中起重要作用,透水性,和镜头透明度。以前的工作表明，特定的蛋白质，如钙调蛋白(CaM)，与AQP0相互作用以调节其透水性；然而，这些研究经常使用AQP0肽，而不是全长蛋白质，探索这些相互作用。此外，几个已知AQP0相互作用伙伴的相互作用的特定区域，即αA和αB-晶状体蛋白，和Pharkinin（CP49）仍然未知。这项研究的目的是使用交联质谱（XL-MS）来鉴定粗晶状体皮质膜部分中与全长AQP0相互作用的蛋白质，并确定相互作用的特定蛋白质区域。我们的研究结果表明,第一次,AQP0N端可以参与蛋白质相互作用。阐明了几种AQP0相互作用伴侣的特定相互作用区域，包括hapkinin，α-晶状体蛋白,连接蛋白-46和连接蛋白-50.此外,两个新的互动伙伴，波形蛋白和连接蛋白-46被鉴定。
