The plasma membrane (PM) acts as the interface between intra- and extracellular environments and exhibits a tightly regulated molecular composition. The composition and amount of PM proteins are regulated by balancing endocytic and exocytic trafficking in a cargo-specific manner, according to the demands of specific cellular states and developmental processes. In plant cells, retrieval of membrane proteins from the PM depends largely on clathrin-mediated endocytosis (CME). However, the mechanisms for sorting PM proteins during CME remain ambiguous. In this study, we identified a homologous pair of ANTH domain-containing proteins, PICALM1a and PICALM1b, as adaptor proteins for CME of the secretory vesicle-associated longin-type R-SNARE VAMP72 group. PICALM1 interacted with the SNARE domain of VAMP72 and clathrin at the PM. The loss of function of PICALM1 resulted in faulty retrieval of VAMP72, whereas general endocytosis was not considerably affected by this mutation. The double mutant of PICALM1 exhibited impaired vegetative development, indicating the requirement of VAMP72 recycling for normal plant growth. In the mammalian system, VAMP7, which is homologous to plant VAMP72, is retrieved from the PM via the interaction with a clathrin adaptor HIV Rev-binding protein in the longin domain during CME, which is not functional in the plant system, whereas retrieval of brevin-type R-SNARE members is dependent on a PICALM1 homolog. These results indicate that ANTH domain-containing proteins have evolved to be recruited distinctly for recycling R-SNARE proteins and are critical to eukaryote physiology.

质膜（PM）充当细胞内和细胞外环境之间的界面，并表现出严格调控的分子组成。根据特定细胞状态和发育过程的需求，通过平衡内吞和胞外运输以货物特定的方式调节PM蛋白的组成和数量。在植物细胞中，从PM中回收膜蛋白主要取决于网格蛋白介导的内吞作用（CME）。但是，在CME期间对PM蛋白进行分选的机制仍然不明确。在这项研究中，我们确定了含有ANTH结构域的一对同源蛋白PICALM1a和PICALM1b，作为分泌小泡相关的浪琴型R-SNARE VAMP72组CME的衔接蛋白。PICALM1在下午与VAMP72的SNARE域和网格蛋白相互作用。PICALM1功能的丧失导致VAMP72的检索错误，而一般的内吞作用不受此突变的影响。的双突变体PICALM1的营养发育受损，表明正常植物生长需要VAMP72回收。在哺乳动物系统中，与植物VAMP72同源的VAMP7通过在CME期间与longin域中的网格蛋白衔接子HIV Rev结合蛋白相互作用而从PM中检索到，而该蛋白在植物系统中不起作用，而brevin型R-SNARE成员依赖于PICALM1同源物。这些结果表明，含有ANTH结构域的蛋白质已经进化为可以循环R-SNARE蛋白质而被明确招募，对于真核生物的生理至关重要。