Bogoeva Vanya Petkova, Petrova Lidiya Plamenova, Trifonov Anton Aleksandrov
Institute of Molecular Biology "Roumen Tsanev", Bulgarian Academy of Sciences, "Acad. G. Bonchev" Str. Bl. 21, 1113, Sofia, Bulgaria.
Sofia University, 5, J. Bourchier Blvd., 1164 Sofia, Bulgaria.
Sci Pharm. 2014 Jun 16;82(4):825-34. doi: 10.3797/scipharm.1404-09. Print 2014 Oct-Dec.
Concanavalin A is a legume lectin which preferentially agglutinates transformed cells and shows antitumor effects on human breast carcinoma cells in vitro and in vivo. It is considered as a new potential antineoplastic agent targeting apoptosis, autophagy, and anti-angiogenesis in preclinical or clinical trials for cancer therapeutics, which has recently become the object of intensive study. In the present investigation, we show the capacity of the lectin to bind manganese, gold, iron, and zinc porphyrins: all potential anticancer agents. The interaction of the legume lectin with the studied compounds has been investigated by tryptophan fluorescence, showing conformational changes within the quaternary and tertiary structures of the protein. The binding of Con A with manganese, gold, and iron porphyrins, as well as adenine, was studied by fluorescence quenching. In contrast, the interaction of Con A with zinc porphyrin caused an increase in Trp fluorescence and a red shift of 10 nm of the emission maximum position. However, the binding of Con A to iron porphyrin was accompanied by a 5 nm blue shift of the emission maximum, and a kD of 0.95 ± 0.13 μM was calculated, respectively. The sigmoidal shape of the curve showed cooperative interactions, which indicated the presence of more than one class of binding site within the Con A molecule for iron porphyrin, confirmed by the Hill slope (h = 1.89±0.46). We have found that the legume lectin interacts with porphyrins and adenine with an affinity (0.14-1.89 µM) similar to that of the non-legume lectin, wheat germ agglutinin. In conclusion, the protein Con A shows new binding activity towards porphyrins with anticancer activities and could find prospective application as a drug delivery molecule that specifically targets cancer cells.
伴刀豆球蛋白A是一种豆类凝集素,它能优先凝集转化细胞,并在体外和体内对人乳腺癌细胞显示出抗肿瘤作用。在癌症治疗的临床前或临床试验中,它被认为是一种靶向凋亡、自噬和抗血管生成的新型潜在抗肿瘤药物,最近已成为深入研究的对象。在本研究中,我们展示了这种凝集素与锰、金、铁和锌卟啉(所有潜在的抗癌剂)结合的能力。通过色氨酸荧光研究了这种豆类凝集素与所研究化合物的相互作用,结果表明蛋白质的四级和三级结构发生了构象变化。通过荧光猝灭研究了伴刀豆球蛋白A与锰、金和铁卟啉以及腺嘌呤的结合。相比之下,伴刀豆球蛋白A与锌卟啉的相互作用导致色氨酸荧光增加,发射最大值位置发生10 nm的红移。然而,伴刀豆球蛋白A与铁卟啉的结合伴随着发射最大值发生5 nm的蓝移,分别计算出解离常数为0.95±0.13 μM。曲线的S形表明存在协同相互作用,这表明伴刀豆球蛋白A分子内存在不止一类铁卟啉结合位点,希尔斜率(h = 1.89±0.46)证实了这一点。我们发现这种豆类凝集素与卟啉和腺嘌呤的亲和力(0.14 - 1.89 µM)与非豆类凝集素麦胚凝集素相似。总之,伴刀豆球蛋白A蛋白对具有抗癌活性的卟啉显示出新的结合活性,有望作为一种特异性靶向癌细胞的药物递送分子得到应用。