PDF(Pubmed)
Abstract：
Palmitoylation is a post-translational lipid modification in which palmitic acid is conjugated predominantly to cysteine residues of target proteins, allowing them to tether to cell membranes. Here, we describe a protocol to perform a stepwise acyl biotin exchange assay to identify protein S-palmitoylation. We describe steps for initial blocking of free thiols in protein lysates, subsequent replacement of thioester-linked palmitate groups with a biotin tag for affinity enrichment, and identification of palmitoylated proteins by SDS-PAGE. For complete details on the use and execution of this protocol, please refer to Leishman et al.1.
摘要：
棕榈酰化是一种翻译后的脂质修饰，其中棕榈酸主要与靶蛋白的半胱氨酸残基缀合。让它们拴在细胞膜上.这里,我们描述了进行逐步酰基生物素交换测定以鉴定蛋白质S-棕榈酰化的方案。我们描述了蛋白质裂解物中游离硫醇的初始阻断步骤，随后用生物素标记替换硫酯连接的棕榈酸酯基团以进行亲和富集，并通过SDS-PAGE鉴定棕榈酰化蛋白。有关此协议的使用和执行的完整详细信息，PleaserefertoLeishmanetal.1.
