Twu J S, Garfinkel A S, Schotz M C
Atherosclerosis. 1975 Nov-Dec;22(3):463-72. doi: 10.1016/0021-9150(75)90025-8.
Rat heart lipoprotein lipase was highly purified by affinity chromatography using heparin-Sepharose 4B. When extracts of acetone powder were applied to columns, lipase activity was firmly bound to the gel matrix and was later eluted with 1.5 M NaCl. At this stage the eluted enzyme was purified 1500-fold. Disc gel electrophoresis yielded a single protein band corresponding with the enzyme activity. The apparent molecular weight was 60,000. The purified enzyme was highly unstable; however, its activity could be partially stabilized by glycerol or ethylene glycol. In studying the purified enzyme we observed: (i) a cofactor in serum was required for full enzymatic activity; ApoLp-Glu could be substituted for this cofactor; (ii) ApoLp-Ser was inhibitory to lipase activity; (iii) NaCl and protamine sulfate also markedly inhibited the lipase activity; (iv) heparin stimulated the enzymatic activity.
使用肝素-琼脂糖4B亲和色谱法对大鼠心脏脂蛋白脂肪酶进行了高度纯化。当将丙酮粉提取物应用于柱时,脂肪酶活性牢固地结合到凝胶基质上,随后用1.5M NaCl洗脱。在此阶段,洗脱的酶被纯化了1500倍。圆盘凝胶电泳产生了一条与酶活性相对应的单一蛋白带。表观分子量为60,000。纯化的酶非常不稳定;然而,其活性可通过甘油或乙二醇部分稳定。在研究纯化的酶时我们观察到:(i)血清中的一种辅因子是酶完全活性所必需的;载脂蛋白Lp-Glu可以替代这种辅因子;(ii)载脂蛋白Lp-Ser抑制脂肪酶活性;(iii)NaCl和硫酸鱼精蛋白也显著抑制脂肪酶活性;(iv)肝素刺激酶活性。