酰基-生物素置换法（Acyl-biotinyl exchange，ABE）是利用生物素取代棕榈酸，进而对棕榈酸修饰的蛋白进行标记检测的方法，是检测S-棕榈酰化蛋白质的常用方法。本研究比较了ABE法中丙酮沉淀与甲醇-氯仿沉淀两种方法对S-棕榈酰化蛋白检测的影响，并对小鼠心脏组织中S-棕榈酰化修饰蛋白进行整体分析，进一步丰富S-棕榈酰化蛋白质的相关研究。首先用N-乙基马来酰亚胺（NEM）对小鼠心脏组织中蛋白质分子上的游离巯基进行封闭，再利用巯基反应性生物素化试剂（HPDP-Biotin）对羟胺（HA）切割后新产生的半胱氨酸巯基进行生物素标记，可检测出小鼠心脏组织中S-棕榈酰化修饰的蛋白。在此反应过程中，均通过蛋白沉淀法除去未反应的NEM、HA及HPDP-Biotin。基于不同沉淀方法对小鼠心脏组织总蛋白中的S-棕榈酰化蛋白进行标记后，利用链霉亲和素修饰琼脂糖珠对已标记上生物素的S-棕榈酰化蛋白进行富集，利用质谱检测分析。本研究共鉴定出50种S-棕榈酰化蛋白，其中丙酮沉淀实验组鉴定出23种S-棕榈酰化蛋白，甲醇-氯仿沉淀实验组鉴定出37种S-棕榈酰化蛋白，两组均鉴定出的蛋白有10种，不同的沉淀方法交叉互补使用可使鉴定结果更加丰富、可靠。

The acyl-biotinyl exchange (ABE) method is a common method for detection of S-palmitoylated proteins by replacing palmitic acid with biotin. In this study, we compared the effects of acetone precipitation and methanol-chloroform precipitation on the detection of S-palmitoylation protein in acyl-biotin exchange method, and analyzed the S-palmitoylated protein in mouse cardiac tissue. First, N-ethylmaleimide (NEM) was used to block the free sulfhydryl on the protein molecules, and then biotinylation reagent (HPDP-Biotin) was used to label the new produced cysteine thiol cleaved by hydroxylamine (HA) in mouse heart tissue. During the ABE reaction, excess unreacted NEM, HA, and HPDP-Biotin were removed by precipitation of the protein. The S-palmitoylated protein in total protein of heart tissue was labeled with ABE reaction based on different precipitation methods, and the S-palmitoylated protein labeled with biotin was enriched by streptavidin agarose beads. The enriched proteins were analyzed by mass spectrometry, and 50 S-palmitoylated proteins were identified. The acetone precipitation assay group identified 23 S-palmitoylated proteins, and the methanol-chloroform precipitation experimental assay group identified 37 S-palmitoylated proteins. 10 proteins were identified in both groups, and 14 newly identified palmitoylated proteins. The result showed that the cross-complementary combination use of different precipitation methods could be helpful for the identification of palmitoylated proteins.