Tsuneoka K, Shikita M, Takatsuki A, Tamura G
J Biochem. 1981 Sep;90(3):611-8. doi: 10.1093/oxfordjournals.jbchem.a133514.
L . P3 cells grown in serum-free synthetic medium produced a colony-stimulating factor (CSF: a sialoglycoprotein stimulating proliferation of granulocyte-macrophage progenitor cells). Addition of tunicamycin (2 microgram/ml) or 2-deoxy-D-glucose (10 mM) to the culture neither decreased the yield of CSF relative to the cell number grown nor induced heterogeneity of the produced CSF. However, CSF produced in the presence of tunicamycin (CSF-tm) was about 23 percent smaller in its molecular weight than normally produced CSF (CSF-normal). Similarly, the addition of 2-deoxy-D-glucose resulted in the formation of a CSF (CSF-dGlc) which was about 16 percent smaller than CSF-normal. However, both CSF-tm and CSF-dGlc seemed to have retained sialic acid residues, because they were focused respectively at pH 4.2 and pH 3.7 upon isoelectric focusing, and both were converted to a pH 5.2 species by treatment with neuraminidase. In addition, CSF-tm was significantly less heat-stable than CSF-normal, whereas CSF-dGlc was only slightly less stable. These results suggest that complete glycosylation of the factor is not necessary for its production nor for its stimulatory action on the proliferation of myeloid stem cells, but is necessary for its maximal stabilization.
在无血清合成培养基中生长的L.P3细胞产生了一种集落刺激因子（CSF：一种刺激粒细胞-巨噬细胞祖细胞增殖的唾液酸糖蛋白）。向培养物中添加衣霉素（2微克/毫升）或2-脱氧-D-葡萄糖（10毫摩尔），相对于生长的细胞数量，既没有降低CSF的产量，也没有诱导所产生的CSF的异质性。然而，在衣霉素存在下产生的CSF（CSF-tm）的分子量比正常产生的CSF（CSF-normal）小约23%。同样，添加2-脱氧-D-葡萄糖导致形成一种CSF（CSF-dGlc），其比CSF-normal小约16%。然而，CSF-tm和CSF-dGlc似乎都保留了唾液酸残基，因为它们在等电聚焦时分别聚焦在pH 4.2和pH 3.7，并且通过用神经氨酸酶处理都转化为pH 5.2的物种。此外，CSF-tm的热稳定性明显低于CSF-normal，而CSF-dGlc的稳定性仅略低。这些结果表明，该因子的完全糖基化对于其产生及其对髓系干细胞增殖的刺激作用不是必需的，但对于其最大程度的稳定是必需的。
