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棕榈酰化修饰 (palmitoylation或S-palmitoylation或S-acylation) according to其连接方式can分forS-棕榈酰化, N-棕榈酰化andO-棕榈酰化. 其中, S-棕榈酰化is指含have16pcs碳原子of饱and棕榈酸andproteinof半胱氨酸残基ofS端结合, 形成不stableof硫酯键, 这种棕榈酰化具have可逆性, 可in时间and空间上调节proteinof功能, 因此棕榈酰化修饰usuallyis指S-棕榈酰化.in所have脂质化修饰中, 棕榈酰化修饰作用most普遍, 影响约20％ofprotein组.
前列腺癌来源ofcell外囊泡富含棕榈酰化protein (Mariscal et al.,2020)
in拟南芥中of棕榈酰化修饰, 促进并定义proteinof膜定位 (Piers et al.,2016)
| Sample Type | 送样量 | 预processandstore |
| 常规动物tissue (脑, 心, 肝, 脾, 肺, 肾, 肌肉, 皮肤etc.) | ≥30 mg | PBSwash去除残留血液and污染物, 迅速剥去脂肪and筋皮etc.结缔tissue, 冲洗干净. 用tissue剪或手术刀将tissue分离成1cm3左右of小块. 液氮速冻, storein-80°C, 干冰transport |
| 软体动物 (血吸虫, 旋毛虫etc.) | ≥200 mg | PBSwash去除来自宿主of污染. 液氮速冻, storein-80°C, 干冰transport |
| 动物软骨tissue | ≥100 mg | PBSwash, 用手术刀将软骨切成1cm3左右of小块. 液氮速冻, storein-80°C, 干冰transport |
| 动物毛发 | ≥50 mg | 用适量2%SDS, 50mM 磷酸钠 (pH7.8) 缓冲液漂洗样品, 去除污染物, 干燥, storein-80°C, 干冰transport |
| 柔软tissue (木本植物of叶, 花etc., 草本科植物, 藻类, 蕨类植物, 大型fungusetc.) | ≥50 mg | 收集样品, 液氮速冻 (such as果样品表面have泥土或明显污染物, 则液氮速冻前用PBS清洗, 吸水纸吸去表面液体) , storein-80°C, 干冰transport |
| 木本科植物of树根, 树皮, 树枝etc. | ≥2 g | 收集样品, 液氮速冻 (such as果样品表面have泥土或明显污染物, 则液氮速冻前用PBS清洗, 吸水纸吸去表面液体) , storein-80°C, 干冰transport |
| 果实, 种子 | ≥200 mg | 收集样品, 液氮速冻 (such as果样品表面have泥土或明显污染物, 则液氮速冻前用PBS清洗, 吸水纸吸去表面液体) , storein-80°C, 干冰transport |
| 果肉 | ≥2 g | 去皮切成1cm3左右小块, 液氮速冻, store于-80°C, 干冰transport |
| 花粉 | ≥10 mg | 植物开花期收集花粉, 解剖显微镜下检查花粉, 用解剖针去除花药碎片etc.杂质, 转入EP管中, 光学显微镜下检查花粉of形态and纯度. store于-80°C, 干冰transport |
| bacteria菌类 | ≥20 mg | 3000g-5000gcentrifuge5-15min, 去上清, PBSwash沉淀三times, 液氮速冻, 菌体-80°Cstore, 干冰transport |
| fungus菌体 | ≥2 g | PBS清洗, 吸水纸吸去表面液体, 液氮速冻, store于-80°C, 干冰transport |
| 悬浮culturecell | ≥1*107 | 1)400g-1000gcentrifuge10minutes收获cell, 弃上清. 2)用预冷ofPBS小心wash片状沉淀物2times, 置于冰上, 去上清. 3)液氮速冻, -80°Cstore, 干冰transport |
| 贴壁culturecell | ≥1*107 | 1)弃掉culture液, 并将culture皿倒置于吸水纸上吸干culture液 2) 加入4°C预冷ofPBS, 平放轻轻摇动1minuteswashcell, 然后弃去PBS. 重复以上操作两times以洗去culture液. 3)将culture皿置于冰上. 向culture皿内加入4°C预冷ofPBS. 用干净ofcell刮棒将cell刮于culture皿of一侧 (动作要快) , 冰上斜置culture皿, 使得缓冲液流向一侧. 移液管吸取溶解产物至预冷ofcentrifuge管内. centrifuge去上清. 4)液氮速冻, -80°Cstore, 干冰transport |
| Serum | ≥50 μL | 收集好of全血室温静置两hours, 3000gcentrifuge10min, 取上清, 液氮速冻, -80°Cstore, 干冰transport |
| 血浆 | ≥50 μL | 收集好of全血加入抗凝剂, 室温静止30minutes, 1300g-2000gcentrifuge10min, 取上清, 液氮速冻, -80°Cstore, 干冰transport |
| 动物乳汁或人乳汁 | ≥1mL | 收集母乳, -80°Cstore, 干冰transport. 若要去除样品中of高丰度酪protein, need超速centrifuge除去. |
| 乳脂肪球膜protein (MFGM) , 即脂质层 | ≥50 μL | 收集母乳, 4度高速centrifuge15min, 取脂质层, 用预冷of5倍体积ofPBSwash3times, 纯水wash一times, -80°Cstore, 干冰transport |
| 脑脊液, 关节液, 淋巴液, 附睾腔液 | ≥20 μL | 1000g-2000gcentrifuge5min, (或using0.22µm滤膜过滤) , 取上清, -80°Cstore, 干冰transport |
| 唾液 | ≥50 μL | 禁食两hours以上, 9-12am取样, 1000g-2000gcentrifuge5min, (或using0.22um滤膜过滤) , 取上清, -80°Cstore, 干冰transport |
| 泪液 | ≥100 μL | 收集样品 (可utilizingcapillary micropipette或Schirmer strip) , 8000-14000g 4度 centrifuge5min, 取上清, -80°Cstore, 干冰transport |
| 尿液 | ≥5 mL | 5000×g 4°C centrifuge30-60min, 取上清, -80°Cstore, 干冰transport |
| 分泌protein-cell分泌液 | ≥2 mL | 10000g-20000gcentrifuge30-60min, (或using0.22um滤膜过滤) , 取上清, -80°Cstore, 干冰transport |
| 分泌protein-脑tissue分泌液 | ≥2 mL | 10000g-20000gcentrifuge30-60min, (或using0.22um滤膜过滤) , 取上清, -80°Cstore, 干冰transport |
| 分泌protein-间质液 | ≥50 μL | 样品收集, -80°Cstore, 干冰transport |
| 外泌体-Serum | ≥2 mL | 收集好of全血室温静置两hours, 3000gcentrifuge10min, 取上清, 液氮速冻, -80°Cstore, 干冰transport |
| 外泌体-Serum | ≥2 mL | 收集好of全血加入抗凝剂, 室温静止30minutes, 1300g-2000gcentrifuge10min, 取上清, 液氮速冻, -80°Cstore, 干冰transport |
| 外泌体-尿液 | ≥30 mL | 5000×g 4°C centrifuge30-60min, 取上清, -80°Cstore, 干冰transport |
| IP样品 | ≥20 μg | 样品洗脱时, recommend采取酸性洗脱模式, wash buffer中不recommend含have去垢剂 (SDS,NP-40,Triton-X100) ,若needusing去垢剂作for洗脱溶液, 则needadoptingSDS-PAGE对样本进行预process, 送样前, 迅速液氮中速冻, 并store于-80°C中, 干冰transport |
1. Neuronal ceroid lipofuscinosis with DNAJC5/CSPα mutation has PPT1 pathology and exhibits aberrant protein palmitoylation. (IF=18.174, Q1, Acta Neuropathologica) DNAJC5 /CSPα
突变of神经元蜡样脂褐质沉积症具havePPT1病理并且表现出异常ofprotein棕榈酰化
2. CD36 facilitates fatty acid uptake by dynamic palmitoylation-regulated endocytosis (IF=-16.6, Q1, Nature Commun)
棕榈酰化protein组揭示CD36of动态棕榈酰化调节脂肪酸of内吞作用
3. Comprehensive palmitoyl-proteomic analysis identifies distinct protein signatures for large and small cancer-derived extracellular vesicles. (IF=14.976, Q1, J Extracell Vesicles)
棕榈酰化protein组综合解析大小不同of癌症来源ofcell外囊泡of独特protein特征
4. ZDHHC18 negatively regulates cGAS-mediated innate immunity through palmitoylation (IF=13.783, Q1, EMBO J)
棕榈酰化protein组揭示cGAS棕榈酰化负调控其enzyme活性as well as先days免疫反应
