01988nas a2200229   4500008004100000022001400041245009800055210006900153260000800222300001000230490000800240520120800248653001701456653003701473653001801510100002001528700001701548700002101565700002601586700002001612856012601632       2021                        eng d  a0022-263100aExpression, purification and crystallization of Asrij, A novel scaffold transmembrane protein0 aExpression purification and crystallization of Asrij A novel sca  cFEB  a65-740 v2543 a<p>Asrij/OCIAD1 is a scaffold transmembrane protein belonging to the Ovarian Carcinoma Immunoreactive Antigen Domain containing protein family. In Drosophila and mouse models, Asrij localizes at the endosomal and mitochondrial membrane and is shown to regulate the stemness of hematopoietic stem cells. Interaction of Asrij with ADP Ribosylation Factor 1 (Arf1) is shown to be crucial for hematopoietic niche function and prohemocyte maintenance. Here, we report the heterologous expression, standardization of detergents and purification methodologies for crystallization of Asrij/OCIAD1. To probe the activity of bacterially expressed Asrij, we developed a protein complementation assay and conclusively show that Asrij and Arf1 physically interact. Further, we find that sophorolipids improve the solubility and monodispersibility of Asrij. Hence, we propose that sophorolipids could be novel additives for stabilization of membrane proteins. To our knowledge, this is the first study detailing methodology for the production and crystallization of a heterologously expressed scaffold membrane protein and will be widely applicable to understand membrane protein structure and function. Graphic</p>
10aAsrij/OCIAD110aMembrane protein crystallography10asophorolipids1 aMotiwala, Zenia1 aDarne, Priti1 aPrabhune, Asmita1 aInamdar, Maneesha, S.1 aKulkarni, Kiran  uhttps://library.ncl.res.in/content/expression-purification-and-crystallization-asrij-novel-scaffold-transmembrane-protein