Purification and characterization of a cytosolic 65-kilodalton phosphoprotein in human leukocytes whose phosphorylation is augmented by stimulation with interleukin 1.
Purification and characterization of a cytosolic 65-kilodalton phosphoprotein in human leukocytes whose phosphorylation is augmented by stimulation with interleukin 1.
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人白细胞中胞质 65 千道尔顿磷蛋白的纯化和表征，其磷酸化通过白细胞介素 1 的刺激而增强。
DOI：
10.1021/bi00410a037
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发表时间：
1988
期刊：
影响因子：
2.9
通讯作者：
Copeland,TD
中科院分区：
文献类型：
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作者：
Matsushima,K;Shiroo,M;Kung,HF;Copeland,TD
[NP]正磷酸盐标记人外周血单核细胞及胞浆的制备用含10%胎牛血清(FCS；HyClone，Logan，VT)的RPMI1640培养液(Advanced Biotech，Inc.，Silver Spring，MD)与10“5M强的松龙(Sigma，St.Louis，MO)在37℃培养约5×106*人PBMC/mLRPMI1640培养液，然后用无磷RPMI1640培养液洗涤两次。此外，在无磷的RPMI1640培养液中，108/毫升的细胞首先在100JICI/毫升[32P]邻苯二甲酸二乙酯的存在下孵育。
Materials and MethodsRadiolabeling of Human PBMC with [nP] Orthophosphate and Preparation of Cytosol. Approximately 5 X 106* human PBMC/mL of RPMI 1640 medium (Advanced Biotechnolo-gies, Inc., Silver Spring, MD) supplemented with 10% fetal calf serum(FCS; Hyclone, Logan, VT) were cultured with10 “5 M prednisolone (Sigma, St. Louis, MO) for 5 h at 37 C. Cells were then pelleted by centrifugation and washed twice with phosphate-free RPMI 1640 medium. Furthermore, 108/mL cells in phosphate-free RPMI 1640 medium were incubated first inthe presence of 100 jiCi/mL [32P] ortho-