Simpson R J, Moritz R L, Van Snick J
Joint Protein Structure Laboratory, Ludwig Institute for Cancer Research, Melbourne Branch, Australia.
Biochem Biophys Res Commun. 1988 Nov 30;157(1):364-72. doi: 10.1016/s0006-291x(88)80056-1.
Murine interleukin 6 (mIL-6) has been synthesized as a fusion protein using a lac operon inducible plasmid in Escherichia coli. The first 8 amino acids are from the N-terminus of bacterial beta-galactosidase and the last 175 amino acids are from residue number 12 to the end of native mIL-6. This fusion protein is equipotent with the native molecule in the hybridoma growth factor assay and has comparable receptor binding characteristics. The two disulfide bridges in mIL-6 have been identified by Staphylococcus aureus V8 protease peptide mapping and Edman degradation of cystine-containing peptides. It has been shown that there are disulfide bonds between Cys46-Cys52 and Cys75-Cys85.
利用大肠杆菌中的乳糖操纵子诱导型质粒，已合成了小鼠白细胞介素6（mIL-6）融合蛋白。前8个氨基酸来自细菌β-半乳糖苷酶的N端，后175个氨基酸来自天然mIL-6第12位残基至末端。在杂交瘤生长因子测定中，这种融合蛋白与天然分子具有同等效力，且具有相似的受体结合特性。通过金黄色葡萄球菌V8蛋白酶肽图谱分析和含胱氨酸肽段的埃德曼降解法，已确定了mIL-6中的两个二硫键。结果表明，在Cys46与Cys52以及Cys75与Cys85之间存在二硫键。
