花椰菜钙调素和兔抗花椰菜钙调的初步应用研究

用花椰菜ＣａＭ和兔抗花椰菜ＣａＭ建立了定量测定了人血清中抗ＣａＭ自身抗体的间接ＥＬＩＳＡ方法。兔抗花椰菜ＣａＭ和兔抗牛脑ＣａＭ对花椰菜ＣａＭ有基本相同的亲和力。用花椰菜ＣａＭ和牛脑ＣａＭ作为检测抗原，测定人血清中抗ＣａＭ自身抗体得到同样的结果。     

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＋     

花椰菜胞外钙调素的性质及促进植物生长作用

花椰菜胞外钙调素的性质及促进植物生长作用刘德龙杨燕生孙大业（中山大学化学系，广州５１０２７５）（河北师范大学生物系）关键词花椰菜，胞外钙调素，花粉萌发，花粉管伸长分类号Ｑ９４２．６钙调素（ＣａＭ）是普遍存在于真核细胞中的一种重要的多功能胞内钙受体，...     

动物体液中的胞外钙调素结合蛋白（快报）

利用以生物素标记钙调素为探针的凝胶覆盖技术检测动物体液中胞外钙调素结合蛋白（ＣａＭＢＰ）。结果，人的唾液中检测到至少３种分子量分别为１４ｋＤ，２４ｋＤ和５２ｋＤ的ＣａＭＢＰｓ。其中，５２ｋＤ蛋白与ＣａＭ的结合依赖于Ｃａ２＋的存在，而２４ｋＤ和１４ｋＤ蛋白则不依赖于Ｃａ２＋。在鸡血清里，检测到以９４ｋＤ，４４／４５ｋＤ蛋白为主的４～５种胞外ＣａＭＢＰｓ，所有的这些蛋白与ＣａＭ的结合都依赖于Ｃａ２－。此外，在牛奶中也检出胞外ＣａＭＢＰｓ。以上结果，证明了在动物中普遍存在胞外ＣａＭＢＰｓ，为胞外钙调素的作用机理提供了新线索。     

小鼠宫腔内用醋酸棉酚的抗生育作用及其与钙调素的关系

于小鼠妊娠第３天每侧子宫角内注入０．１８ｍｇ醋棉酚混悬液（０．０５ｍＬ），对照组注入等量溶剂，结果表明，醋酸棉酚对小鼠有显著的抗生育作用，抗生育率为９３％，抗着床率为７７％，钙调素（ＣａＭ）含量的测定表明，实验组子宫组织的ＣａＭ含量明显低于对照，证明子宫内ＣａＭ的降低是局部用棉酚抗生育作用的机理之一。     

高迁移组染色体蛋白-17的分离提取及其抗体ELISA测定方法的建立

纯化高迁移组染色体蛋白－１７（ＨＭＧ－１７）,建立检测抗ＨＭＧ－１７自身抗体的ＥＬＩＳＡ方法,探讨抗ＨＭＧ－１７抗体与临床疾病的关系。用水煮沸法提取,甲酸－吡啶缓冲液酸化和ＳｅｐｈａｄｅｘＧ－１５０凝胶过滤技术,自小牛胸腺组织中分离ＨＭＧ－１７,用此纯化物包被,方阵滴定,建立测定抗ＨＭＧ－１７抗体的间接ＥＬＩＳＡ方法,并进行方法学考核和临床标本检测。纯化的ＨＭＧ－１７抗原经十二烷基硫酸钠－聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）分析,呈现一条分子量约９２００的蛋白带;建立的ＥＬＩＳＡ方法批内平均变异系数（ＣＶ）为５．８％,批间ＣＶ值为９．６％,ＥＬＩＳＡ抑制试验最高抑制率达８７．２％;临床检测结果表明,在系统性红斑狼疮（ＳＬＥ）、混合性结缔组织病（ＭＣＴＤ）和干燥综合征（ＳＳ）患者中分别有１７．４％～３７．３％的抗ＨＭＧ－１７抗体的阳性检出率（Ｐ＜０．０１或Ｐ＜０．０５）,而在其他自身免疫病和内科疾病者中,抗ＨＭＧ－１７抗体均为低水平。抗ＨＭＧ－１７抗体ＥＬＩＳＡ测定法具有较好的特异性和重复性,便于常规检测,该类抗体的存在似与某些自身免疫病（尤其是ＳＬＥ）的关系较为密切     

对流免疫电泳试验在肝片吸虫病诊断中的应用

目的探讨对流免疫电泳试验在肝片吸虫病诊断中的应用。方法用对流免疫电泳试验（CIET）检测肝片吸虫患者、肝片吸虫病兔、正常兔、血吸虫病兔和血吸虫患者血清的抗体。结果以CIET检测肝片吸虫患者和病兔血清中的抗体，阳性率分别为90．3％和92．3％。同时检测正常兔、血吸虫病兔和血吸虫病人血清，皆无沉淀线形成，阴性反应。结论CIET用于肝片吸虫病临床诊断具有较好的特异性。     

重组人钙调素的表达及其McAb的制备与精子CaM定位

用基因重组技术构建人钙调素基因Ⅲ（ｈＣａＭⅢｃＤＮＡ）表达载体ｐＢＶ／ｈＣａＭⅢ,并在大肠杆菌ＤＨ５α中经热诱导获得可溶性ＣａＭ蛋白的高效表达．将纯化的重组ｈＣａＭ与异型双功能剂ＳＰＤＰ及鼠血清白蛋白（ＭＳＡ）交联,免疫Ｂａｌｂ／ｃ小鼠,用常规细胞融合技术制备单克隆抗体（ＭｃＡｂ）,得到３株稳定分泌单克隆抗体的杂交瘤细胞．间接ＥＬＩＳＡ和斑点免疫结果证实,三种单克隆抗体均与自制的ｒｈＣａＭ和ＣａＭ标准品起特异反应．用免疫组化法对精子中ＣａＭ定位,发现ＣａＭ主要分布于精子头颈部,不育组ＣａＭ＋精子率（（４５．０±７．５）％）显著低于生育组（（６８．５±１０．５）％）．     

花椰菜天然钙调素抗体的特性及其ELISA定量法的建立

钙调素是一种具有多种调节功能的钙结合蛋白质，其免疫原性很弱，本文采用两次基础免疫和多次加强免疫的方法，获得了免疫花椰菜天然钙调素抗血清，用免疫双扩散法鉴定，其效价为１：３２钙调节不易与固相载体结合，我们先用０．２％戊二醛处理聚苯乙烯板载体，再用于包被钙调素，在上述基础上建立了定量测定植物钙调素的竞争性酶联免疫吸附测定（ＥＬＩＳＡ）技术及检测动物血清中抗钙调素抗体的间接ＥＬＩＳＡ技术。     

钙调素参与植物线粒体琥珀酸脱氢酶（SDH）活性调节的研究

对纯化的玉米线粒体ＳＤＨ进行ＣａＭ含量测定电泳分析，对ＮＡＤＫ的激活及外源ＣａＭ对ＳＤＨ活性的影响研究，结果表明：ＳＤＨ中可能含ＣａＭ亚基，并且ＳＤＨ活性可能受Ｃａ（２＋）·ＣａＭ调节．     

Fluorescence Spectra Studies on the Interaction between Lanthanides and Calmodulin

ＩｎｔｒｏｄｕｃｔｉｏｎＬａｎｔｈａｎｉｄｅ (Ｌｎ3 )ｃａｎ ｐｒｏｍｏｔｅｔｈｅｇｒｏｗｔｈｏｆｃｒｏｐｓａｎｄｉｎｃｒｅａｓｅｔｈｅｉｒｐｒｏｄｕｃｔｉｏｎ .Ｈｏｗｅｖｅｒ ,Ｌｎ3 ｃａｎａｌｓｏａｆｆｅｃｔａｎｉｍａｌｔｉｓｓｕｅｓａｎｄｈａｓｓｏｍｅｔｏｘｉｃｆｕｎｃｔｉｏｎｓａｔｃｅｒｔａｉｎｃｏｎｃｅｎｔｒａｔｉｏｎｓ[1] .ＴｈｅｍｅｃｈａｎｉｓｍｂｙｗｈｉｃｈＬｎ3 ｅｘｅｒｔｓｂｉｏｌｏｇｉｃａｌｅｆｆｅｃｔｓｒｅｍａｉｎｓｕｎｃｌｅａｒ.ＳｔｕｄｉｅｓｓｈｏｗｔｈａｔＬｎ3 ｃａｎｉ…     

白芷细胞外CaM结合位点的扫描电镜定位研究

利用胶体金标记的具有生物活性的钙调素（Ｃａｌｍｏｄｕｌｉｎ,ＣａＭ）,对白芷愈伤组织培养细胞胞外钙调素结合位点进行了扫描电镜定位,发现白芷愈伤组织培养细胞表面有不同密度的金颗粒,证明其细胞表面存在着ＣａＭ结合位点．     

东海原甲藻钙调蛋白的分离纯化及鉴定

以东海原甲藻(Prorocentrum donghaiense Lu)为材料,经热变性、离心处理,上清液依次经DEAE纤维素阴离子交换层析和苯基-Sepharose CL- 4B疏水层析分离纯化钙调蛋白(CaM).SDS-PAGE和Western blot检测结果显示:东海原甲藻中CaM的相对分子质量为16 ku.采用该实验方法首次有效地从海洋浮游植物东海原甲藻中分离纯化出钙调蛋白.     

Calmodulin bifurcates the local Ca2+ signal that modulates P/Q-type Ca2+ channels

Acute modulation of P/Q-type (alpha1A) calcium channels by neuronal activity-dependent changes in intracellular Ca2+ concentration may contribute to short-term synaptic plasticity, potentially enriching the neurocomputational capabilities of the brain. An unconventional mechanism for such channel modulation has been proposed in which calmodulin (CaM) may exert two opposing effects on individual channels, initially promoting ('facilitation') and then inhibiting ('inactivation') channel opening. Here we report that such dual regulation arises from surprising Ca2+-transduction capabilities of CaM. First, although facilitation and inactivation are two competing processes, both require Ca2+-CaM binding to a single 'IQ-like' domain on the carboxy tail of alpha1A; a previously identified 'CBD' CaM-binding site has no detectable role. Second, expression of a CaM mutant with impairment of all four of its Ca2+-binding sites (CaM1234) eliminates both forms of modulation. This result confirms that CaM is the Ca2+ sensor for channel regulation, and indicates that CaM may associate with the channel even before local Ca2+ concentration rises. Finally, the bifunctional capability of CaM arises from bifurcation of Ca2+ signalling by the lobes of CaM: Ca2+ binding to the amino-terminal lobe selectively initiates channel inactivation, whereas Ca2+ sensing by the carboxy-terminal lobe induces facilitation. Such lobe-specific detection provides a compact means to decode local Ca2+ signals in two ways, and to separately initiate distinct actions on a single molecular complex.     

Calmodulin modulation of ion channels and receptors

Ion channels and receptors are the structural basis for neural signaling and transmission. Recently, the function of ion channels and receptors has been demonstrated to be modulated by many intracellular and extracellular chemicals and signaling molecules. Increasing evidence indicates that the complexity and plasticity of the function of central nervous system is determined by the modulation of ion channels and receptors. Among various mechanisms, Ca 2+ signaling pathways play important roles in neuronal activity and some pathological changes. Ca 2+ influx through ion channels and receptors can modulate its further influx in a feedback way or modulate other ion channels and receptors. The common feature of the modulation is that Ca 2+ /calmodulin (CaM) is the universal mediator. CaM maintains the coordination among ion channels/receptors and intracellular Ca 2+ homeostasis by feedback modulation of ion channels/receptors activity. This review focuses on the modulating processes of ion channels and receptors mediated by CaM, and further elucidates the mechanisms of Ca 2+ signaling.     

Calmodulin modulation of ion channels and receptors?

：Ion channels and receptors are the structural basis for neural signaling and transmission. Recently, the function of ion channels and receptors has been demonstrated to be modulated by many intracellular and extracellular chemicals and signaling molecules. Increasing evidence indicates that the complexity and plasticity of the function of central nervous system is determined by the modulation of ion channels and receptors. Among various mechanisms, Ca 2+ signaling pathways play important roles in neuronal activity and some pathological changes. Ca 2+ influx through ion channels and receptors can modulate its further influx in a feedback way or modulate other ion channels and receptors. The common feature of the modulation is that Ca 2+ /calmodulin (CaM) is the universal mediator. CaM maintains the coordination among ion channels/receptors and intracellular Ca 2+ homeostasis by feedback modulation of ion channels/receptors activity. This review focuses on the modulating processes of ion channels and receptors mediated by CaM, and further elucidates the mechanisms of Ca 2+ signaling.     

Effects of extracellular calmodulin on pollen germination and tube growth

The effects of calmoddin (CaM) antagonist W7-agarose, anti-CaM serum and exogenous purified CaM on pollen germination and tube growth ofForsythia suspensu were studied. The pollen germination and tube growth were inhibited or completely stopped by CaM antagonist W7-agarose. The addition of exogenous purified CaM stimulated pollen germination and tube growth, whereas the same amount of bovine serum albumin (BSA) had no effect. The inhibitory effects caused by W7-agarose and anti-CaM serum could be reversed completely by the addition of exogenous purified CaM. The tube growth of germinated pollen was also inhibited or completely stopped by W7-agarose. The results suggest that endogenous extracellular CaM initiates pollen germination and tube growth, whereas exogenous CaM enhances the above processes.