Phosphorylation of the multidrug resistance associated protein gene encoded protein P190.
Phosphorylation of the multidrug resistance associated protein gene encoded protein P190.
复制标题
多药耐药相关蛋白基因编码蛋白 P190 的磷酸化。
DOI：
10.1021/bi00010a024
复制
发表时间：
1995
期刊：
影响因子：
2.9
通讯作者：
Center,MS
中科院分区：
文献类型：
--
作者：
Ma,L;Krishnamachary,N;Center,MS
材料和方法材料。[32P]正磷酸（8500-9000 Ci1 mmol）购自Dupont/New England Nuclear。Staurosporine来自Behring Diagnostics， l-（5-异喹啉基磺酰基）-2-甲基哌嗪（H-7）来自Sigma。Chelerythrine来自LC Services Corp.。分离出的对阿德里亚霉素耐药的HL60细胞按先前描述的方法制备（Marsh et al., 1986）。这些细胞对选择剂的抵抗力增加了80倍。如前所述，获得了HL60/ADR的自发逆转物和指定的HL60/ADR/R （Krishnamachary et al., 1994）。对阿霉素的抗性增加了3倍。含PI90推断序列肽抗体的制备。由Research Genetics， Hunstville， al合成了序列为QRGLFYSMAKDAGLV的肽，该肽对应于MRP末端c末端的推断序列（Cole et al., 1992）。抗血清指定为6KQ，并根据该肽制备（Krishnamachary et al., 1994）。
MATERIALS AND METHODSMaterials.[32P] Orthophosphoric acid (8500—9000 Ci1 mmol) was purchased from Dupont/New England Nuclear. Staurosporine was from Behring Diagnostics, and l-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7) was from Sigma. Chelerythrine was obtained from LC Services Corp. Cell Lines. HL60 cells isolated for resistance to adria-mycin were prepared as described previously (Marsh et al., 1986). These cells exhibit an 80-fold increase in resistance to the selecting agent. A spontaneous revertant of HL60/ADR and designated HL60/ADR/R was obtained as previously described (Krishnamachary et al., 1994). The revertant exhibits about a 3-fold increase in resistance to adriamycin. Preparation of an Antibody against a Peptide Containing the PI90 Deduced Sequence. A peptide having the sequence QRGLFYSMAKDAGLV and corresponding to the deduced sequence of the extreme C-terminal end of MRP (Cole et al., 1992) was synthesized by Research Genetics, Hunstville, AL. Antiserum designated 6KQ and prepared against this peptide was as described previously (Krishnamachary et al., 1994).