一种新的平滑肌调控蛋白Calponin

Calponin为一种平滑肌特有的调控蛋白, 分子克隆的证据表明, 它具有两种亚型, α型和β型分别由292和252个氨基酸组成它能与肌动蛋白结合, 抑制肌球蛋白ATP酶活性和平滑肌收缩其与肌动蛋白结合域有38个氨基酸残基(第145～182位), 丝氨酸175在调宁蛋白与肌动蛋白的相互作用中起重要作用, 它还能与钙调蛋白结合, 呈钙依赖性, 其结构域在第52～144位残基调宁蛋白的机能受磷酸化与脱磷酸化的调节.     

调宁蛋白T184的定点突变、表达、纯化和鉴定

为增强调宁蛋白对平滑肌收缩的抑制作用,用PCR重叠延长法使调宁蛋白基因中编码第184位苏氨酸的ACT突变为编码丙氨酸的GCC,将此PCR的突变产物装入到质粒载体pAED₄后,转化至E.coli BL₂₁(DE₃)中, 构建的重组转化子用酶切和测序鉴定.诱导含有重组转化子的E.coli获得高效表达,经SDS-PAGE和蛋白质印迹鉴定后,采用反复冻融法及葡聚糖凝胶层析柱分离,初步纯化出调宁蛋白突变体T184A.     

植物渗调蛋白的研究进展

植物在逆境下会产生许多逆境响应蛋白,渗调蛋白（Osmotins)是其中重要的一种,该蛋白是一种逆境适应,蛋白,其基因的表达受到干旱,盐渍,病原侵染,乙烯,ABA等因子的诱导,与植物的抗旱,耐盐和抗病性等有关。作者对渗调蛋白的存在,特性,功能和基因调控规律作以下简要综述。     

碱性抗菌蛋白的研究进展

抗菌蛋白和抗菌肽在非特划性免疫中起到越来越重要的作用,是解决多药性抗药菌和细菌毒素引起的难治性感染症的有希望的方法,本文就hCAP18抗菌蛋白的结构、功能、作用机制及应用作一综述。     

原核生物中的类钙调蛋白

钙是重要的生命元素之一,真核生物中普遍存在介导钙信号的钙调蛋白已有深入的研究,证明钙调蛋白是细胞复杂调控系统中的一个重要成员,但是在原核生物中是否也存在类似的蛋白因子却一直说法不一.自80年代初在大肠杆菌(Escherichia coli)中首次发现类钙调蛋白(calmodulin-like-protein)以来,已在多种原核生物中陆续发现了类钙调蛋白的存在,证明其可能参与了原核生物的孢子形成,细胞分裂,生物固氮,异型胞形成和兰细菌光合作用等多种调控功能.文章综述了近年来这一领域的部分研究成果.     

平滑肌22α蛋白在血管稳态和血管重构中的作用

有关血管稳态和重构的分子机制一直是近年来的研究热点,也被视为治疗血管损伤性疾病的突破点.大量研究证实,血管损伤修复及病理性重构过程与血管平滑肌细胞(vascular smooth muscle cells,VSMCs)的表型转化、异常增殖与迁移、细胞衰老关系密切.平滑肌22α(smooth muscle 22α,SM2...     

豌豆钙调蛋白基因的克隆及七种来源钙调蛋白基因的分析

从豌豆（Ｐｉｓｕｍ ｓａｔｉｖｕｍ Ｌ．）中提取总ＲＮＡ, 逆转录出ｃＤＮＡ 第一条链后, 以大麦钙调蛋白结构基因两端的寡聚核苷酸为引物, 用ＰＣＲ方法合成豌豆钙调蛋白基因, 克隆到Ｂｌｕｅ－ｓｃｒｉｐｔ 载体上并测定其全序列。结果与已知的苜蓿（Ｍｅｄｉｃａｇｏ ｓａｔｉｖａ Ｌ．）、水稻（Ｏｒｙｚａ ｓａｔｉｖａＬ．）、大麦（Ｈｏｒｄｅｕｍ ｖｕｌｇａｒｅ Ｌ．）、电鳗（Ｅｌｅｃｔｒｏｐｈｏｒｉｄａｅ）、根瘤（Ａｓｐｅｒｇｉｌｌｕｓ ｎｉｄｕｌａｎｓ）、酵母（Ｓａｃｃｈａｒｏｍｙｃｅｓｃｅｒｅｖｉｓｉａｅ） 钙调蛋白基因相比,同源性较高（９１．３％ —６０．８％ ）,其不同部分则常常是Ｃ和Ｔ相替换。进一步分析发现,上述七种来源的钙调蛋白基因之间,常有某些核苷酸替换方式占优势,它们对于氨基酸密码子的使用也各有偏爱     

Calponin in Non-Muscle Cells

Calponin is an actin filament-associated regulatory protein expressed in smooth muscle and non-muscle cells. Calponin is an inhibitor of the actin-activated myosin ATPase. Three isoforms of calponin have been found in the vertebrates. Whereas the role of calponin in regulating smooth muscle contractility has been extensively investigated, the function and regulation of calponin in non-muscle cells is much less understood. Based on recent progresses in the field, this review focuses on the studies of calponin in non-muscle cells, especially its regulation by cytoskeleton tension and function in cell motility. The ongoing research has demonstrated that calponin plays a regulatory role in non-muscle cell motility. Therefore, non-muscle calponin is an attractive target for the control of cell proliferation, migration and phagocytosis, and the treatment of cancer metastasis.     

心脏特异表达Calponin 1转基因小鼠的心脏功能分析

目的建立心脏特异表达Calponin 1转基因小鼠,研究Calponin 1对心脏发育及心肌病的调节作用。方法利用心脏特异启动子α-MHC构建转基因表达载体,显微注射法建立Calponin 1转基因小鼠,PCR法鉴定转基因小鼠的基因型,Western Blot检测Calponin 1在心脏组织中的表达,心脏超声检测转基因小鼠的心脏结构和功能,HE染色和Masson染色检测转基因小鼠心脏的病理改变。结果 Calponin 1在野生型小鼠心脏中有表达,在扩张型心肌病小鼠的心脏组织表达降低。通过显微注射法,建立了2个心脏组织Calponin 1基因高表达的转基因小鼠系。与野生型小鼠相比,Calponin 1转基因小鼠收缩期左室内径（LVID,systolic）增加28%（P〈0.01,n=12）,舒张期左室内径（LVID,diastolic）增加16.2%（P〈0.01,n=12）,收缩期左室后壁厚度（LVPW,systolic）减小15.7%（P〈0.01,n=12）,舒张期左室后壁厚度（LVPW,diastolic）减小21%（P〈0.01,n=12）,射血分数（ejection fraction,EF）降低11.5%（P〈0.01,n=12）,短轴内径缩短率（fraction shortening,FS）降低14.6%（P〈0.05,n=12）。转基因小鼠心脏组织病理H＆E染色和Masson染色显示,转基因小鼠心室扩张,心肌细胞不均匀肥大,细胞间隙变大,心肌间质纤维增多。结论 Calponin 1在心脏特异过表达引起转基因小鼠心脏左室内径增加,收缩期容积和舒张期容积显著增大,心室壁变薄,射血分数及短轴缩短率降低等扩张性心肌病表型,推测Calponin 1是参与心肌病病理发生的基因之一。     

血管损伤的新靶点：平滑肌22 alpha

血管平滑肌细胞(vascular smooth muscle cell,VSMC)表型转化是血管损伤性疾病动脉粥样硬化、高血压和血管成形术后再狭窄等的共同病理生理过程.平滑肌22 alpha (smooth muscle 22 alpha, SM22α) 是一种VSMC分化标志物,其表达具有平滑肌组织特异性和细胞表型特异性. 该蛋白不仅作为一种肌动蛋白细胞骨架相关蛋白参与VSMC骨架组构和收缩调节,它还参与VSMC的增殖、炎症和氧化应激等进程. 本文就SM22α 的结构特征及其在VSMC血管损伤中的作用机制进行综述.     

旋转加压种植法在新型生物人工复合血管体外内皮化过程中的实验研究

目的：探讨在新型生物人工复合血管内腔面联合种植平滑肌细胞和内皮细胞的方法,比较研究旋转加压种植与普通灌注种植两种方法的内皮化效果,方法：先制备新型生物人工复合血管及获取培养鉴定平滑肌细胞和内皮细胞,再和旋转加压种植与普通灌注种植两种方法将平滑肌细胞和内皮细胞培养种植于新型复合血管内腔面,以光镜及扫描电镜等观察评价内皮化的效果。结果：旋转加压种植2小时末的复合血管腔内有大量内皮细胞,旋转加压种植9天后已形成完整的内皮细胞单层;普通灌注种植的复合血管内腔有内皮细胞附着,分布不均匀,未形成完整的内皮细胞层,结论：以旋转加压种植法在新型复合血管内腔面联合种植平滑肌细胞和内皮细胞效果满意,基本实现内皮化,可以满足复合血管内皮化的要求。     

Calponin control of cerebrovascular reactivity: therapeutic implications in brain trauma

Calponin (Cp) is an actin-binding protein first characterized in chicken gizzard smooth muscle (SM). This review discusses the role of Cp in mediating SM contraction, the biochemical process by which Cp facilitates SM contraction and the function of Cp in the brain. Recent work on the role of Cp in pathological states with emphasis on traumatic brain injury is also discussed. Based on past and present data, the case is presented for targeting Cp for novel genetic and pharmacological therapies aimed at improving outcome following traumatic brain injury (TBI).     

Store-operated Ca entry and depolarization explain the anomalous behaviour of myometrial SR: Effects of SERCA inhibition on electrical activity,Ca and force

In the myometrium SR Ca²⁺ depletion promotes an increase in force but unlike several other smooth muscles, there is no Ca²⁺ sparks-STOCs coupling mechanism to explain this. Given the importance of the control of contractility for successful parturition, we have examined, in pregnant rat myometrium, the effects of SR Ca²⁺-ATPase (SERCA) inhibition on the temporal relationship between action potentials, Ca²⁺ transients and force. Simultaneous recording of electrical activity, calcium and force showed that SERCA inhibition, by cyclopiazonic acid (CPA 20 μM), caused time-dependent changes in excitability, most noticeably depolarization and elevations of baseline [Ca²⁺]_i and force. At the onset of these changes there was a prolongation of the bursts of action potentials and a corresponding series of Ca²⁺ spikes, which increased the amplitude and duration of contractions. As the rise of baseline Ca²⁺ and depolarization continued a point was reached when electrical and Ca²⁺ spikes and phasic contractions ceased, and a maintained, tonic force and Ca²⁺ was produced. Lanthanum, a non-selective blocker of store-operated Ca²⁺ entry, but not the L-type Ca²⁺ channel blocker nifedipine (1–10 μM), could abolish the maintained force and calcium. Application of the agonist, carbachol, produced similar effects to CPA, i.e. depolarization, elevation of force and calcium. A brief, high concentration of carbachol, to cause SR Ca²⁺ depletion without eliciting receptor-operated channel opening, also produced these results. The data obtained suggest that in pregnant rats SR Ca²⁺ release is coupled to marked Ca²⁺ entry, via store operated Ca²⁺ channels, leading to depolarization and enhanced electrical and mechanical activity.