Broxterman H J, Smit J S, van der Plas J, van Langevelde A, Belfroid R D, van Kempen G T, van der Krogt J A
Cancer Lett. 1985 Sep 15;28(2):177-86. doi: 10.1016/0304-3835(85)90073-4.
Binding of [3H]flunitrazepam and [3H]spiperone to membrane preparations isolated by high speed centrifugation of hamster, rabbit and human melanoma cell homogenates was analyzed. All melanoma cell types expressed a high density of specific binding sites for [3H]flunitrazepam (3-4 pmol/mg protein) with a high affinity (Kd about 30 nM). This binding was independent of melanin content of cells and could be classified, based on competition experiments, as a Ro 5-4864-like binding type. Specific [3H]spiperone binding to these cell lines clearly revealed at least two types of binding sites: a low affinity, high capacity type of binding site (Kd greater than 100 nM, Bmax about 50 pmol/mg protein) and a high affinity, low capacity binding site (Kd less than 1 nm, Bmax 30 fmol/mg protein). Binding of spiperone to the low affinity, high capacity site appeared displaceable by NM 113 and dependent on melanin content of the cells and probably represents binding to melanin. Analysis of drug binding to melanoma membrane cell preparations and correlation with drug effects should include the possible involvement of binding to melanin.
分析了用仓鼠、兔子和人类黑色素瘤细胞匀浆高速离心分离得到的膜制剂与[3H]氟硝西泮和[3H]螺哌隆的结合情况。所有黑色素瘤细胞类型均表达高密度的[3H]氟硝西泮特异性结合位点（3 - 4 pmol/mg蛋白质），且亲和力高（Kd约为30 nM）。这种结合与细胞的黑色素含量无关，根据竞争实验，可归类为Ro 5 - 4864样结合类型。[3H]螺哌隆与这些细胞系的特异性结合清楚地显示至少有两种结合位点：一种是低亲和力、高容量的结合位点（Kd大于100 nM，Bmax约为50 pmol/mg蛋白质）和一种高亲和力、低容量的结合位点（Kd小于1 nM，Bmax为30 fmol/mg蛋白质）。螺哌隆与低亲和力、高容量位点的结合似乎可被NM 113取代，且依赖于细胞的黑色素含量，可能代表与黑色素的结合。对黑色素瘤细胞膜制剂的药物结合分析以及与药物效应的相关性分析应包括与黑色素结合的可能影响。
