Functional groups at the catalytic site of F1 adenosine triphosphatase

The protection of F1 ATPase by inorganic phosphate, ADP, ATP, and magnesium ion against inactivation by 1-fluoro-2,4-dinitrobenzene, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole, and 1-(ethoxycarbonyl)-2-ethoxy-1,2-dihydroquinoline, respectively, has been investigated. Dissociation equilibrium constants and rate constants for the labeling reactions have been deduced from a quantitative treatment of the kinetic data. Comparison of these dissociation constants with each other and with the corresponding literature values indicates that the essential Tyr, Arg, Lys, and Glu or Asp residues are indeed located at the catalytic site of the enzyme. Examination of the rate constants for the labeling reactions in the presence of excess inorganic phosphate, ADP, ATP, or magnesium ion, respectively, suggests that the essential phenol and amino groups are located nearer to the bound inorganic phosphate or the gamma-phosphate group than to the alpha- or beta-phosphate group of the bound ATP, that the essential guanidinium group is located nearer to the alpha- or beta-phosphate group than to the gamma-phosphate group of the bound ATP or the bound inorganic phosphate, and that the essential carboxylate group is located slightly farther away but complexed with magnesium ion which it shares with the bound inorganic phosphate. A mechanism consistent with these topographical relationships is proposed for the catalytic hydrolysis and synthesis of ATP.     

Studies of the mechanisms for the induction of in vivo tumor immunity. VI. Induction of specific and nonspecific cell-mediated immunity in tumor-bearing hosts and its correlation with transplantation tumor immunity

Studies were performed to determine the development of cell-mediated cytotoxic response at tumor site in C57BL/6 mice bearing progressively growing FBL-3 ascites leukemia. The effectors isolated from tumor ascites are found to be highly cytotoxic for leukemic target cells. The levels of cytotoxicity obtained with effectors isolated from tumor site are generally higher than those obtained with immune mice. This cytotoxicity is both specific and nonspecific. The specific cytotoxicity against tumor-associated antigen is mainly mediated by T cells and the nonspecific cytotoxicity against unrelated tumor cells is mediated largely by macrophages. The T-cell-enriched preparation did not give significant natural killer activity. When testing the ability of these effectors to produce in vivo immunity against the challenge of FBL-3, it was found that only T cells could confer the transplantation-type immunity, but the immunity was transient. The macrophage-enriched preparation isolated from tumor ascites failed to give in vivo protection. These findings indicate that in FBL-3 system, mice with progressively growing tumors are able to develop immune response against tumor cells. However, this immunity is probably interfered with by a suppressor factor(s) or suppressor cells which restrict their activity to eliminate the tumor cells effectively.     

动物内源褪黑素调控生物节律的研究进展

内源褪黑素对人类和其他哺乳动物的节律行为具有调控功能。生物节律是自然进化赋予生命的基本特征之一,生物体的生命活动受到生物节律的控制与影响。在哺乳动物中,节律调控中心是松果体,其主要功能是合成和分泌褪黑素。褪黑素广泛参与生物体节律行为的调节,本文从褪黑素的产生和作用机制,分别阐述褪黑素对昼夜节律行为和多种年节律行为的调控作用,同时明确褪黑素与生物钟及神经内分泌系统的直接作用和反馈互动的复杂集合,进一步揭示褪黑素调控生物节律的重要作用,以期为褪黑素的基础研究以及未来探究生物体的生物钟内源性发生机制提供参考。     

茄子果实相关农艺性状全基因组关联分析研究进展与展望

茄子是重要的园艺作物,也是茄科植物中种植最广泛的蔬菜之一。茄子果实相关农艺性状是一种复杂的数量性状,传统育种选育效率低、周期长。高通量测序技术与生物信息学技术的快速发展,使得全基因组关联分析(genome-wide association study, GWAS)在解析茄子果实相关复杂农艺性状的遗传规律方面展现出巨大的应用前景。本文对全基因组关联分析在茄子的果形、果色等果实相关农艺性状中的研究进展进行了综述;针对茄子数量性状遗传研究中普遍存在的“丢失遗传力”(missing heritability)问题,从4个GWAS策略在茄子果实相关农艺性状研究中的应用热点出发,提出了未来茄子GWAS的发展对策;并结合当前茄子遗传改良的实践需求,展望了GWAS策略在茄子分子育种领域的广阔应用前景。本文为今后利用GWAS解析各种茄子果实相关性状的遗传基础以及选育符合消费者需求的果实材料提供了理论依据和参考。     

苹果柠檬酸合酶3基因家族成员鉴定及表达分析

柠檬酸合酶(citrate synthase 3, CS3)是细胞代谢途径中的关键酶之一,其活性调节着生物体的物质和能量代谢过程。本研究旨在从苹果全基因组中鉴定CS3基因家族成员,并进行生物信息学和表达模式分析,为研究苹果CS3基因的潜在功能提供理论基础。利用BLASTp基于GDR数据库鉴定苹果CS3家族成员,通过Pfam、SMART、MEGA5.0、clustalx.exe、ExPASy Proteomics Server、MEGAX、SOPMA、MEME和WoLF PSORT等软件分析CS3蛋白序列基本信息、亚细胞定位情况、结构域组成、系统进化关系以及染色体定位情况。利用酸含量的测定和实时荧光定量PCR (real-time fluorescence quantitative polymerase chain reaction, qRT-PCR)技术检测苹果6个CS3的组织表达和诱导表达特性。苹果CS3基因家族包含6个成员,这些CS3蛋白包括473−608个不等的氨基酸残基,等电点分布在7.21−8.82。亚细胞定位结果显示CS3蛋白分别定位在线粒体和叶绿体。系统进化分析可将其分为3类,各亚家族基因数量分别为2个。染色体定位结果显示,CS3基因分布在苹果不同的染色体上。蛋白二级结构以a-螺旋为主,其次是无规则卷曲,b-转角所占比例最小。筛选的6个家族成员在不同苹果组织中均有表达,整体表达趋势从高到低依次为MdCS3.4相对表达含量最高,MdCS3.6次之,其他家族成员相对表达量依次为MdCS3.3>MdCS3.2>MdCS3.1>MdCS3.5。qRT-PCR结果显示,MdCS3.1和MdCS3.3基因在酸含量较低的‘成纪1号’果肉中相对表达量最高,酸含量较高的‘艾斯达’果肉中MdCS3.2和MdCS3.3基因相对表达量最高。因此,本研究对不同苹果品种中CS3基因相对表达量进行了检测,并分析了其在苹果果实酸合成过程中的作用。结果表明,CS3基因在不同苹果品种中的相对表达量存在差异,为后续研究苹果品质形成机制提供了参考。     

斑马鱼急性无机砷暴露后肝脏差异表达基因的生物信息学分析

砷是一种致癌物,是心血管、外周血管疾病、神经疾病、糖尿病和各种癌症的致病因素。目的:利用GO数据库和KEGG数据库等生物信息学方法对GEO数据库数据中的差异表达基因进行评价。利用生物信息学分析软件对差异基因进行功能富集、功能注释分析和生存分析。利用Cytoscape上的蛋白-蛋白相互作用网络(Protein-protein interaction network, PPI)软件对179个差异基因进行筛选和分析。结果发现126个基因作用于蛋白靶点,其中有10个基因为关键基因分别为:PSMB3、HSP701、HSPE1、STIP1、HSPD1、HSP70、DNAJB1B、HSP90AA1.1、HSPA9H和TCP1。核心基因主要作用于内质网中的蛋白质加工通路。这可能会为砷对肝脏损伤的潜在生物标志物和生物学机制提供新的思路。     

PLZF-RARα融合基因的生物学功能研究

构建了8个PLZF-RARα融合基因突变体.用“滞后”胶实验证实PLZF-RARα与PML-RARα一样,亦能以同二聚体的形式结合到维甲酸反应元件(RARE)上,且PLZF的POZ结构域介导PLZF-RARα同二聚体的形成和稳定.但两者与RARE的结合类型存在差异,在DR5G,PML-RARα的结合强度大于PLZF-RARα;而在DR5T,则是PLZF-RARα强于PML-RARα.进一步工作证实PLZF-RARα能与RXR形成异二聚体,并产生4种复合物.用免疫沉淀法发现PLZF-RARα亦能与PLZF形成异二聚体,而且也是通过POZ结构域介导PLZF-RARα和PLZF异二聚体的形成.同PML-RARα一样,PLZF-RARα对RARE的结合反应亦受维甲酸调控.     

左旋千金藤啶碱的非经典安定剂特性——电生理和行为的研究

在记录神经元放电频率的实验中,发现麻醉或麻痹大鼠给予DA受体拮抗剂SPD,使VTA,SNC和DA神经元放电频率增加.但随着剂量的增加,SPD选择性地使VTA的DA神经元放电完全抑制,可被DA受体激动剂APO所翻转.提示这种抑制作用可能是DI.但SPD不抑制SNCDA神经元放电.在计数自发放电的神经元数目的实验中,观察到:慢性给予SPD21d后,大鼠VTA部位自发放电的DA神经元数目减少,有量效关系.可被APO所翻转.相反,同样给药并不影响SNC部位自发放电的DA神经元数目.当急性给予SPD时,VTA自发放电的DA神经元数目也减少,而SNC自发放电的DA细胞数不受影响.在行为实验中,SPD能抑制APO引起的定型活动,但SPD本身引起的僵住症很弱,仅能维持15min,并且无剂量依赖关系.上述结果提示:SPD选择性抑制VTADA神经元,而对SNCDA神经元的抑制作用很弱,引起的锥体外系副作用弱,可能发展成为新一代的非经典安定剂.