Department of Early Discovery Biochemistry, Genentech, Inc., South San Francisco, CA 94080, USA.
FEBS Lett. 2011 Aug 4;585(15):2501-6. doi: 10.1016/j.febslet.2011.06.033. Epub 2011 Jul 2.
Palmitoylation of the Wnt and Hedgehog proteins is critical for maintaining their physiological functions. To date, there are no reported studies that characterize the cellular distribution of the palmitoylated forms of these proteins. Here, we describe the subcellular localization of palmitoylated Wnt and Sonic Hedgehog by using a highly sensitive and non-radioactive labeling method that utilizes alkynyl palmitic acid. We show that palmitoylated Wnt and Sonic Hedgehog localize to cellular membrane fractions only, highlighting a role for palmitoylation in the membrane association of these proteins. The method described herein has the utility to validate inhibitors of Wnt and Hedgehog acyltransferases in drug discovery, and enables further investigations of the role of palmitoylation in the secretion and signaling of these proteins.
蛋白质棕榈酰化对于维持 Wnt 和 Hedgehog 蛋白的生理功能至关重要。迄今为止，尚未有研究报道这些蛋白的棕榈酰化形式的细胞分布特征。在这里，我们描述了使用高度敏感且非放射性的炔基棕榈酸标记方法，对棕榈酰化 Wnt 和 Sonic Hedgehog 的亚细胞定位。我们发现棕榈酰化 Wnt 和 Sonic Hedgehog 仅定位于细胞膜部分，这突出了棕榈酰化在这些蛋白与膜结合中的作用。本文所描述的方法可用于验证 Wnt 和 Hedgehog 酰基转移酶抑制剂在药物发现中的应用，并能够进一步研究棕榈酰化在这些蛋白分泌和信号转导中的作用。
