Photoaffinity labeling of P-glycoprotein in multidrug resistant cells with photoactive analogs of colchicine.
Photoaffinity labeling of P-glycoprotein in multidrug resistant cells with photoactive analogs of colchicine.
复制标题
用秋水仙碱的光活性类似物对多药耐药细胞中的 P-糖蛋白进行光亲和标记。
DOI：
10.1016/0006-291x(89)90830-9
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发表时间：
1989
影响因子：
3.1
通讯作者：
Agresti,M
中科院分区：
文献类型：
--
作者：
Safa,AR;Mehta,ND;Agresti,M
合成了两种秋水仙碱的光活性放射性标记类似物，N-(对叠氮基[3, 5-[3 H]苯甲酰基)氨基己酰脱乙酰秋水仙碱([3 H] NABC])和N-(对叠氮基-[3-125 I]水杨基)氨基己酰脱乙酰秋水仙碱([125 I] NASAC)，并用于鉴定膜囊泡中的秋水仙碱特异性受体。多重耐药（MDR）变体DC-3F/VCRd-5L中国仓鼠肺细胞。 [3 H] NABC 和 [125 I] NASC 均特异性地光标记 DC-3F/VCRd-5L 细胞膜囊泡中显着的 150-180 kDa 多肽。光标记的多肽通过 MDR 相关 P-糖蛋白 (P-gp) 特异性的单克隆抗体 C219 进行免疫沉淀，表明该蛋白与 P-gp 的同一性。 1000 μM 的秋水仙碱可将 P-gp 的 [3 H] NABC 光标记降低 72%。此外，100 μM 秋水仙碱、长春新碱、长春花碱、阿霉素和放线菌素 D 分别抑制 [125 I]NASC 光标记 45%、88.8%、91.1%、61.5% 和 51%。然而，甲氨蝶呤不影响P-gp的[125I]NASC光标记，表明P-gp秋水仙碱受体对这些细胞耐药的药物具有多药特异性。
Two photoactive radiolabeled analogs of colchicine, N-(p-azido [3, 5-[3 H] benzoyl) aminohexanoyldeacetylcolchicine ([3 H] NABC]) and N-(p-azido-[3-125 I] salicyl) aminohexanoyldeacetylcolchicine ([125 I] NASC) were synthesized and used to identify colchicine-specific acceptor (s) in membrane vesicles from multidrug resistant (MDR) variant DC-3F/VCRd-5L Chinese hamster lung cells. Both [3 H] NABC and [125 I] NASC specifically photolabeled a prominent 150–180 kDa polypeptide in membrane vesicles from DC-3F/VCRd-5L cells. The photolabeled polypeptide was immunoprecipitated by monoclonal antibody C219 specific for the MDR-related P-glycoprotein (P-gp) indicating the identity of this protein with P-gp. Colchicine at 1000 μM reduced [3 H] NABC photolabeling of P-gp by 72%. Furthermore, 100 μM of colchicine, vincristine, vinblastine, doxorubicin and actinomycin D inhibited [125 I] NASC photolabeling by 45, 88.8, 91.1, 61.5, and 51% respectively. However, methotrexate did not affect the [125 I] NASC photolabeling of P-gp, indicating the multidrug specificity of the P-gp colchicine acceptor for drugs to which these cells are resistant.