摘要:
The methods and compositions disclosed herein concern the synthesis of a novel class of “two-headed” phospholipid-phosphoinositide hybrids possessing a carbon backbone, such as 2,3-diacylthreitol, erythritol or a synthetic module. The second phospholipid head group allows introduction of a biochemical or chemical moiety in a position orthogonal in space to those occupied by the phosphoinositide head group and the two acyl chains. The diacyl moieties allow for the incorporation of Pea-PIP2 into a lipid bilayer, while the Ptdlns(4,5)P2 moiety in the aqueous layer is specifically recognized by lipid binding proteins. In alternative embodiments of the invention, reporters, for example biotin, fluorophores and/or spin labels, are attached to the free amino group of the head groups of such molecules to specifically target the reporters to the lipid-water interface.
摘要翻译:本文公开的方法和组合物涉及具有碳骨架的新型“双头”磷脂 - 磷酸肌醇混合物的合成,例如2,3-二酰基硫代糖,赤藓糖醇或合成模块。 第二个磷脂头组允许在与磷酸肌醇头基和两个酰基链占据的位置上在空间正交的位置引入生物化学或化学部分。 二酰基部分允许将Pea-PIP 2 N 2掺入脂质双层中,而水层中的Ptdlns(4,5)P 2 H 2部分被 脂质结合蛋白。 在本发明的替代实施方案中,记录者例如生物素,荧光团和/或旋转标记连接到这些分子的头基的游离氨基上,以将记录特异性地靶向脂质 - 水界面。
摘要:
A lipid assay kit for detection of a lipid kinase or phosphatase and method of use thereof. The assay is preferably a competitive assay wherein the product lipid has a stronger affinity to the lipid detector protein than the substrate lipid. The lipid recognition motif is preferably a pleckstrin homology (PH) domain.
摘要:
Lipid assay method, kit, and apparatus involving exposure of a protein, having a lipid recognition motif that interacts with a target lipid and a competing lipid, to a solution containing the competing lipid, and determining whether the target lipid is present in the solution. The target lipid has a stronger affinity to the lipid recognition motif than does the competing lipid. The lipid recognition motif is preferably a pleckstrin homology (PH) domain, with the target lipid being a phosphoinositide. The assay determines activity of a lipid kinase, the target lipid being a phosphorylation product of a reaction between the lipid kinase and a substrate lipid. The assay can be a cancer screening method for detection of cancer cells, where detection of certain levels of a PI(3,4,5)P3 target lipid is an indicator of a cancer cell.