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在全细胞溶解酸中,可溶性和膜性甘氨酸蛋白的甘氨酸结节概况
Jesús S Aguilar Díaz de León1, Jorvani Cruz Villarreal1, Erandi P Kapuruge1
1School of Molecular Sciences and the Biodesign Institute - Center for Personalized Diagnostics, Arizona State University, P.O. Box 876401, Tempe, AZ, 85287, USA.
Analytical biochemistry
|September 12, 2023
概括
甘氨酸结核分析 (GNA) 现在分析细胞解质中的膜糖蛋白,揭示癌细胞的独特甘氨酸配置文件,并验证甘氨酸转移酶活性监测.
科学领域:
- 格莱科米克斯 (Glycomics) 是一个有趣的药物.
- 癌症生物学 癌症生物学
- 生物化学 生物化学
背景情况:
- 糖结分析 (GNA) 是一种用于量化单糖单位的糖化学技术.
- 此前,GNA已被应用于血/血清,用于癌症检测和预测.
- 从培养细胞中获得的膜糖蛋白以前是GNA无法获得的.
研究的目的:
- 适应GNA用于在培养细胞中分析膜糖蛋白.
- 为了比较肝细胞癌 (HepG2) 细胞与白血病细胞 (THP-1,K562) 和正常的PBMCs的甘氨酸样本.
- 为了证明GNA能够检测糖系转移酶活性变化的能力.
主要方法:
- 细胞溶解物的甲醇/形式沉.
- 液相甲基化 (LPP) 用于增强的糖结分析.
- 糖甘结合特异性单糖体单位的相对量化.
主要成果:
- 通过使用LPP,GNA成功适应了膜糖蛋白,改善了信号对噪声,产量和精度.
- 与白血病细胞和PBMCs相比,肝细胞癌 (HepG2) 细胞表现出较高的天线化,N-甘氨酸分支和α2,6-化.
- 白血病细胞表现出高水平的糖脂相关的糖节点比HepG2细胞和PBMCs.
- 在HepG2细胞中抑制fucosyltransferase降低了天线fucosylation,证实了GNA对糖系转移酶活性的敏感性.
结论:
- 现在GNA适用于培养细胞中的膜糖蛋白,扩大其在糖化学研究中的实用性.
- 独特的甘氨酸特征区分癌细胞类型和正常细胞.
- GNA提供了一种灵敏的方法来监测甘氨酸转移酶活性及其对甘氨酸表达的影响.
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