Mohamad Saharuddin Bin, Nagasawa Hideko, Uto Yoshihiro, Hori Hitoshi
Department of Biological Science and Technology, Faculty of Engineering, University of Tokushima, Minamijyosanjimacho-2, Tokushima, 770-8506 Japan.
Anticancer Res. 2002 Nov-Dec;22(6C):4297-300.
Gc protein has been reported to be a precursor of Gc protein-derived macrophage activation factor (GcMAF) in the inflammation-primed macrophage activation cascade. An inducible beta-galactosidase of B cells and neuraminidase of T cells convert Gc protein to GcMAF.
Gc protein from human serum was purified using 25(OH)D3 affinity column chromatography and modified to GcMAF using immobilized glycosidases (beta-galactosidase and neuraminidase) The sugar moiety structure of GcMAF was characterized by lectin blotting by Helix pomatia agglutinin. The biological activities of GcMAF were evaluated by a superoxide generation assay and a phagocytosis assay.
We successfully purified Gc protein from human serum. GcMAF was detected by lectin blotting and showed a high biological activity.
Our results support the importance of the terminal N-acetylgalactosamine moiety in the GcMAF-mediated macrophage activation cascade, and the existence of constitutive GcMAF in human serum. These preliminary data are important for designing small molecular GcMAF mimics.
据报道，在炎症引发的巨噬细胞激活级联反应中，Gc蛋白是Gc蛋白衍生的巨噬细胞激活因子（GcMAF）的前体。B细胞的诱导型β-半乳糖苷酶和T细胞的神经氨酸酶将Gc蛋白转化为GcMAF。
用人血清中的Gc蛋白通过25(OH)D3亲和柱色谱法进行纯化，并用固定化糖苷酶（β-半乳糖苷酶和神经氨酸酶）将其修饰为GcMAF。用滨螺凝集素通过凝集素印迹法对GcMAF的糖部分结构进行表征。通过超氧化物生成试验和吞噬试验评估GcMAF的生物学活性。
我们成功从人血清中纯化出Gc蛋白。通过凝集素印迹法检测到GcMAF，并显示出高生物学活性。
我们的结果支持了末端N-乙酰半乳糖胺部分在GcMAF介导的巨噬细胞激活级联反应中的重要性，以及人血清中组成型GcMAF的存在。这些初步数据对于设计小分子GcMAF模拟物很重要。
