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胎牛血清的熱滅活內(nèi)毒素污染影響人類T淋巴細(xì)胞蛋白質(zhì)組和磷酸化2016-10-11 14:35
目前的研究提供了新的信息關(guān)于FCS熱失活的影響和改變FCS-LPS濃度在細(xì)胞蛋白表達(dá),并在人類T淋巴母翻譯修飾。熱失活和有限合伙人污染FCS的調(diào)節(jié)蛋白質(zhì)的表達(dá)和磷酸化參與基本細(xì)胞功能,如蛋白質(zhì)合成、細(xì)胞骨架穩(wěn)定性、氧化應(yīng)激調(diào)控和細(xì)胞凋亡。因此,研究強(qiáng)調(diào)需要考慮熱失活和有限合伙人FCS的污染因素可以影響T淋巴母細(xì)胞蛋白質(zhì)組。 英文原文: Fetal calf serum heat inactivation and lipopolysaccharide contamination influence the human T lymphoblast proteome and phosphoproteome Background The effects of fetal calf serum (FCS) heat inactivation and bacterial lipopolysaccharide (LPS) contamination on cell physiology have been studied, but their effect on the proteome of cultured cells has yet to be described. This study was undertaken to investigate the effects of heat inactivation of FCS and LPS contamination on the human T lymphoblast proteome. Human T lymphoblastic leukaemia (CCRF-CEM) cells were grown in FCS, either non-heated, or heat inactivated, having low (< 1 EU/mL) or regular (< 30 EU/mL) LPS concentrations. Protein lysates were resolved by 2-DE followed by phospho-specific and silver nitrate staining. Differentially regulated spots were identified by nano LC ESI Q-TOF MS/MS analysis. Results A total of four proteins (EIF3M, PRS7, PSB4, and SNAPA) were up-regulated when CCRF-CEM cells were grown in media supplemented with heat inactivated FCS (HE) as compared to cells grown in media with non-heated FCS (NHE). Six proteins (TCPD, ACTA, NACA, TCTP, ACTB, and ICLN) displayed a differential phosphorylation pattern between the NHE and HE groups. Compared to the low concentration LPS group, regular levels of LPS resulted in the up-regulation of three proteins (SYBF, QCR1, and SUCB1). Conclusion |