Anees M, Benbow E W
Division of Biochemistry and Molecular Biology, School of Biological Sciences, University of Manchester, UK.
J Enzyme Inhib. 1996;10(3):195-201. doi: 10.3109/14756369609030313.
Dansyl fluoride (Dan-F), an active site directed fluorescent inhibitor of guanidinobenzoatase (GB), has been used for the location of tumour cells in frozen sections of human squamous cell carcinoma and colonic carcinoma tissues. The tumour cell surfaces having active GB bind Dan-F and fluoresce blue. The surrounding normal epithelial lung cell surfaces fail to bind Dan-F and hence lack fluorescence, whilst the normal colon cell surfaces have another isoenzymic form of GB, bind Dan-F and fluoresce blue. Kinetic studies have shown that Dan-F is an irreversible inhibitor of GB, and Dan-GB complexes are not dissociated with SDS and high salt concentration. However hydroxylamine (1 M) can dissociate Dan-GB complexes in the presence of 0.1% SDS, both on membrane-bound and in free solution. These studies suggest that Dan-F is a potent inhibitor of GB, and in very low concentration (3 x 10(-8) M) can be used as a novel fluorescent probe for the location of tumour cells in histological sections of human tissues.
丹磺酰氟(Dan-F)是一种针对胍基苯甲酸酶(GB)活性位点的荧光抑制剂,已被用于在人鳞状细胞癌和结肠癌组织的冰冻切片中定位肿瘤细胞。具有活性GB的肿瘤细胞表面结合Dan-F并发出蓝色荧光。周围正常的肺上皮细胞表面未能结合Dan-F,因此缺乏荧光,而正常结肠细胞表面有GB的另一种同工酶形式,结合Dan-F并发出蓝色荧光。动力学研究表明,Dan-F是GB的不可逆抑制剂,Dan-GB复合物不会被SDS和高盐浓度解离。然而,在0.1% SDS存在的情况下,羟胺(1 M)可以在膜结合和游离溶液中解离Dan-GB复合物。这些研究表明,Dan-F是GB的有效抑制剂,在极低浓度(3×10⁻⁸ M)下可作为一种新型荧光探针用于在人体组织组织切片中定位肿瘤细胞。