Gc蛋白衍生的巨噬细胞激活因子（GcMAF）的制备及其结构表征与生物学活性
Preparation of Gc protein-derived macrophage activating factor (GcMAF) and its structural characterization and biological activities.
作者信息
Mohamad Saharuddin Bin, Nagasawa Hideko, Uto Yoshihiro, Hori Hitoshi
机构信息
Department of Biological Science and Technology, Faculty of Engineering, University of Tokushima, Minamijyosanjimacho-2, Tokushima, 770-8506 Japan.
出版信息
Anticancer Res. 2002 Nov-Dec;22(6C):4297-300.
BACKGROUND
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.
MATERIALS AND METHODS
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.
RESULTS
We successfully purified Gc protein from human serum. GcMAF was detected by lectin blotting and showed a high biological activity.
CONCLUSION
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.
背景
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。
材料与方法
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。
结果
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。
结论
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。
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