植物钙调素的分离纯化及其特性研究

本文以烟草愈伤组织和花椰菜这花序为材料，分离纯化钙调素。所得ＣａＭ的纯度用ＳＤＳ－ＰＡＧＥ鉴定；生物活性用其对牛心肌环腺苷酸磷二酯酶（ＰＤＥ）的激活作用测定（ＣａＭ）的产量用酶联免疫吸附法（ＥＬＩＳＡ）测定。并比较了上Ｐｈｎｅｙｌ－ＳｅｐｈａｒｏｓｅＣＬ－４Ｂ柱前对ＣａＭ溶液的不同处理。纯化的烟草愈伤组织和菜花ＣａＭ，具有牛脑ＣａＭ所特有的一些性质。它对牛心肌ＰＤＥ有明显激活作用，在含有Ｃａ＾２＋     

动物脑组织中钙调蛋白的分离纯化

本文主要以动物脑组织为材料,经热变性、离心、葡聚糖SephadexG-50分离纯化,通过紫外吸收图谱、二步酶解法检验钙调蛋白的纯度、浓度。结果显示,利用本法可分离纯化得到的钙调蛋白对PDE的激活倍数为83.3,比标准的钙调蛋白(对PDE的激活倍数为67.4)更具生物活性.     

Structure and function of a calmodulin-dependent smooth muscle myosin light chain kinase

Summary In smooth muscle the M_r 20,000 light chain of myosin is phosphorylated by a calmodulin-dependent protein kinase. It consists of 2 subunits: calmodulin, an acidic protein of M_r 17,000 that binds 4 moles of Ca²⁺; and a larger protein of M_r circa 130,000. Activation of the kinase is dependent upon their association in the presence of Ca²⁺. Cyclic AMP-dependent protein kinase phosphorylation of the myosin light chain kinase occurs at 2 sites. It decreases the affinity of the kinase for calmodulin and a reduction in the rate of light chain phosphorylation occurs. The kinase has an overall asymmetric shape composed of a globular head and tail region for the skeletal muscle enzyme. Trypsin digestion of this kinase releases a fragment of M_r 36,000 from the globular region that contains the catalytic and calmodulin binding sites. Chymotrypsin digestion of the kinase from smooth muscle generates a fragment of M_r 80,000 that does not contain the calmodulin binding or cyclic AMP-dependent protein kinase phosphorylation sites. It is a Ca²⁺-independent form of the kinase that phosphorylates the light chain of myosin. These structural features indicate a regulatory role for the kinase in smooth muscle phosphorylation and contraction.     

Localization of calmodulin and calmodulin-like protein and their functions in biomineralization in P. fucata

Calmodulin (CaM) and calmodulin-like protein (CaLP) are two proteins involved in biomineralization. Their localizations in Pinctada fucata mantle epithelia were studied by Western blot (WB) analysis of the nuclear/cytosol fraction of primary cultured Pinctada fucata mantle cells and immunogold electron microscopy. The results showed a completely different distribution of these two proteins at the subcellular level. CaM was distributed throughout both the nucleus and cytoplasm of the mantle epithelium but CaLP was distributed only in the cytoplasm. The functions of these two proteins in biomineralization were investigated by shell regeneration. During this process, the expressions of CaM and CaLP were greatly enhanced in different organelles of the mantle epithelium. Overexpression of these two proteins and a mutant of calmodulin-like protein (M-CaLP) that lacks an extra C-terminal tail in MC3T3-E1 promoted the mRNA expression of osteopontin, a biomineralization marker for osteoblasts. All of the results indicated that CaM and CaLP have completely different distributions in the mantle epithelium and affect the biomineralization process at different levels. The extra C-terminal tail of CaLP is important for its functions in biomineralization in Pinctada fucata.     

钙调素特异性拮抗剂(TFP)对大鼠着床的影响

在大鼠孕４ｄ宫腔内一次性注入钙调素特异性拮抗剂－三氟拉嗪,每侧子宫角０．０５ｍＬ,具有显著的抗着床作用,抗着床率高达１００％,而注入等容量生理盐水的对照组为０％。     

蚯蚓钙调素结合蛋白的研究

以赤子爱胜蚓(Eisenia Foetida)为材料,通过DEAE-Fast Flow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。     

Interaction of smooth muscle relaxant drugs with calmodulin and cyclic nucleotide phosphodiesterase

Summary Some smooth muscle relaxant drugs with an unknown mechanism of action have been tested for their interaction with calmodulin and with calmodulin-induced cyclic nucleotide phosphodiesterase (PDE) activity. The affinity of these drugs for calmodulin does not parallel their inhibitory effect on the calmodulin activation of PDE. The lack of parallelism could be due to a binding of the drugs to different sites on calmodulin; furthermore a binding of papaverine, octylonium bromide and felodipine to PDE molecule, might also be considered to explain their inhibitory effect on PDE basal activity. The myolytic effect of octylonium bromide and pinaverium bromide may be due to their interaction with calmodulin-dependent systems.     

番茄碱对棉铃虫毒性分析

本文主要研究了番茄碱对棉铃虫的毒性作用．实验结果表明：（1）用不同浓度的番茄碱（tomatine）浸泡的棉花叶饲喂棉铃虫时，随浓度的增加及时间的延长，对棉铃虫的杀伤力增强．结果显示：饲喂第三天时，1mmol／L的浓度引起48％死亡，2mmol／L时60％死亡，4mmol／L时76％死亡，其IC50为1．78mmol／L．（2）用紫外光谱分析测得虫体内钙调蛋白的含量，结果显示：tomatine对其体内钙调蛋白的影响较大，随tomatine浓度的增加，钙调蛋白的含量逐渐降低．（3）紫外光谱图谱显示：番茄碱与钙调蛋白形成了一种新的物质（其特征吸收峰在190nm-220nm）．说明tomatine对棉铃虫的杀伤作用与钙调蛋白有关，此项研究为进一步探讨tomatine的作用分子机制提供了科学参数．     

钙调素拮抗剂对荷瘤小鼠主要器官钙调素（CaM）水平的影响

本文采用磷酸二酯酶（ＰＤＥ）法,测定了小檗胺衍生物ＥＢＢ对荷Ｓ１８０瘤小鼠的主要器官和瘤体ＣａＭ含量的影响。实验结果表明,ＥＢＢ具有使荷瘤后小鼠脑、肺、肝、脾、肾五个主要器官ＣａＭ由不正常趋向正常的能力,而已知抗肿瘤药物丝裂霉素和环磷酰胺单独使用时此种能力较弱,但这两种抗肿瘤药物与ＥＢＢ联合作用时,能使荷瘤鼠诸器官的ＣａＭ水平趋向正常。结果提示,专一性强的ＣａＭ拮抗剂在抑制肿瘤的同时可能还具有维护