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蚯蚓钙调素结合蛋白的研究
Studies on CaM binding Proteins from Earthworm
【摘要】 以赤子爱胜蚓(EiseniaFoetida)为材料,通过DEAE-FastFlow离子交换层析、CaM-Sepharose亲和层析,分离纯化得到蚯蚓钙调素结合蛋白(CaMBPs)。纯化的CaMBPs对CaM激活的环核苷酸磷酸二酯酶活性有抑制作用,而且这种抑制作用可通过加入过量的CaM达到完全恢复。SDS-PAGE显示CaMBPs有3条明显主带,在EGTA存在时表现分子量分别为62,49和30kD。紫外扫描测定含量分别为7.17%,7.31%和51.8%。用生物素-CaM覆盖法检测到3种CaM结合蛋白,与SDS-PAGE结果一致。酶活性测定实验表明在蚯蚓CaMBPs中有Ca2+-ATPase活性,但无NAD激酶活性。
【Abstract】 Calmodulin binding proteins were purified from earthworm (Eisenia Foetida) by a procedure involving DEAE Fast Flow,CaM Sepharose 4B chromatographies.The purified CaM binding proteins inhibited the activity of CaM dependent PDE and the degree of inhibition increased with augmentation of the CaMBPs.But this inhibition could be eliminated by adding an excess of earthworm CaM.This appeared to be the typical character of CaM binding Proteins.Three main bands of purified CaMBPs were shown on SDS PAGE with apparent molecular weight of 62,49 and 30kD.Their content percent was determined as 7.17%,7.31% and 51.8% by gel scaning respectively.Three kinds of CaMBPs were detected by biotinylated CaM overlay method.The assay of enzyme activity indicates that there was CaM dependent Ca 2+ ATPase activity,but no NAD kinase one.
【Key words】 Calmodulin binding protein; Earthworm; Calmodulin; CaM Sepharose 4B affinity column;
- 【文献出处】 北京大学学报(自然科学版) ,ACTA SCICENTIARUM NATURALUM UNIVERSITIS PEKINESIS , 编辑部邮箱 ,1997年06期
- 【分类号】Q51
- 【被引频次】18
- 【下载频次】142