MiaB protein from Thermotoga maritima. Characterization of an extremely thermophilic tRNA-methylthiotransferase

In Escherichia coli, the MiaB protein catalyzes the methylthiolation of N-6-isopentenyl adenosine in tRNAs, the last reaction step during biosynthesis of 2-methylthio-N-6-isopentenyl adenosine (ms2i6A-37). For the first time the thermophilic bacterium Thermotoga maritima is shown here to contain such a MiaB tRNA-modifying enzyme, named MiaBTm, and to synthesize ms2i6A-37 as demonstrated by an analysis of modified nucleosides from tRNA hydrolysates. The corresponding gene (TM0653) was identified by sequence similarity to the miaB gene cloned and expressed in E. coli. MiaBTm was purified to homogeneity and thoroughly characterized by biochemical and spectroscopic methods. It is a monomer of 443 residues with a molecular mass of 50,710 kilodaltons. Its amino acid sequence shares the CysXXX-CysXXCys sequence with MiaB from E. coli as well as with biotin synthase and lipoate synthase. This sequence was shown to be essential for chelation of an iron-sulfur center and for activity in these enzymes. As isolated, MiaBTm contains both iron and sulfide and an apoprotein form can coordinate up to 4 iron and 4 sulfur atoms per polypeptide chain. UV-visible absorption, resonance Raman, variable temperature magnetic circular dichroism, and EPR spectroscopy of MiaBTm indicate the presence of a [4Fe-4S]+2/+1 cluster under reducing and anaerobic conditions, whereas [3Fe-4S]+1 and [2Fe-2S]+2 forms are generated under aerobic conditions. The redox potential of the [4Fe-4S]+2/+1 transition is -495 +/- 10 mV (versus the normal hydrogen electrode). Finally, the expression of MiaBTm from T. maritima in an E. coli mutant strain lacking functional miaB gene allowed production of ms2i6A-37. These results provide further information on the enzymes involved in methylthiolation of tRNAs.     

黄瓜幼苗光合作用对高温胁迫的响应与适应

以‘津优35号’黄瓜幼苗为试材,研究高温(HT: 42 ℃/32 ℃)和亚高温(SHT: 35 ℃/25 ℃)胁迫对黄瓜幼苗光合作用及生长量的影响.结果表明: 高温、亚高温明显抑制幼苗生长.随着胁迫时间的延长,黄瓜幼苗叶片的光合速率(P_n)逐渐降低,胞间CO₂浓度(C_i)趋于升高,气孔导度(g_s)、蒸腾速率(T_r)、光呼吸速率(P_r)和暗呼吸速率(D_r)先上升后下降,高温、亚高温引起P_n降低的主要原因是非气孔限制.高温、亚高温可使黄瓜幼苗叶片的暗下光系统Ⅱ最大光化学效率(F_v/F_m)、光下实际光化学效率(Φ_PSII)、光化学猝灭系数(q_P)和电子传递效率(ETR)显著降低,初始荧光(F_o)和非化学猝灭系数(NPQ)逐渐升高.随着胁迫时间的延长,HT处理的RuBP羧化酶(RuBPCase)和Rubisco活化酶(RCA)活性及其mRNA表达量逐渐降低,而SHT处理的胁迫初期变化不大,3 d后趋于降低;HT和SHT处理的景天庚酮糖-1,7-二磷酸酶(SBPase)和果糖-1,6-二磷酸醛缩酶(FBA)活性与mRNA表达均呈先升高后降低趋势.可见,适宜光强下短时亚高温处理黄瓜幼苗不会产生明显光抑制,高温胁迫会对其PSⅡ反应中心造成严重损伤;光合酶受高温胁迫诱导,但其诱导效应与温度升高幅度和高温持续时间有关.     

日光温室黄瓜发育过程中主要芳香物质和脂肪酸含量的变化

以'新泰密刺'为试验材料,比较分析了日光温室条件下,不同月份黄瓜商品成熟果实及其发育过程中的主要芳香物质及亚麻酸、亚油酸含量差异.结果显示:(1)黄瓜果实中挥发性物质主要有2E,6Z-壬二烯醛、2E-壬烯醛、2E-己烯醛、2,6-壬二烯醇、正己醛、戊醛、2-戊烯醛和3,6-壬二烯醇.其中2E,6Z-壬二烯醛相对含量以5月份最高,1月份其次,11月份最低;具有黄瓜香气的2,6-壬二烯醇和3,6-壬二烯醇,以及与黄瓜风味品质密切相关的2E,6Z-壬二烯醛/2E-壬烯醛比值以1月份最高,5月份其次,11月份最低.(2)1月份黄瓜果实的亚麻酸和亚油酸含量显著大于5月份,说明低温弱光有利于不饱和脂肪酸的形成和积累.(3)随着果实发育天数的增加,黄瓜2E,6Z-壬二烯醛、2,6-壬二烯醇和3,6-壬二烯醇相对含量,2E,6Z-壬二烯醛/2E-壬烯醛比值均较大幅度升高,商品成熟期达到高峰,若延迟采收,上述指标多趋于下降;2E-壬烯醛、2E-己烯醛、正己醛、戊醛等的相对含量多随果实发育而降低,商品成熟期2E-壬烯醛、2E-己烯醛和正己醛降到最低,之后快速升高.研究表明,果实发育过程中亚麻酸和亚油酸含量及二者的比例与2E,6Z-壬二烯醛和2E-壬烯醛相对含量及其比值的变化规律基本相似,说明黄瓜风味品质与其亚麻酸和亚油酸含量有一定相关性.     

间充质干细胞分化为心肌细胞的诱导方法研究进展

间充质干细胞作为一种取材方便、易于分离培养、体外扩增快、免疫原性低的成体干细胞,具有自我更新和多向分化潜能,可在体内外不同的诱导条件下分化为心肌细胞,是理想的心肌再生治疗的种子细胞。本文综述了间充质干细胞分化为心肌细胞的诱导方法,包括化学试剂、中药制剂、机械力和电磁刺激、心肌环境因子、损伤组织条件培养、组织工程方法等,为其在心肌损伤性疾病尤其是心肌梗死治疗中的应用提供基础。     

根黄酮在植物生长和营养中的作用

 本文简要综述了80年代以来国际上对根黄酮在调节植物根生长、完善根功能、影响氮素循环及在施加他感作用方面的研究进展,并对有关方面作了一些展望,以期引起植物营养工作者对植物次生物质的注意。     

猕猴桃不同季节的嫁接法比较试验

在湖北的试验表明,猕猴桃春季嫁接以切接法较好,夏秋季嫁接则以单芽枝腹接法较好;嫁接后采用遮阳网遮荫可以显著提高嫁接成活率,但对出圃率无明显影响。     

具有一般形式饱和接触率SEIS模型的周期解

利用重合度的延拓定理,导出了具有一般形式饱和接触率SEIS模型周期解的存在性准则.     

Rosin Surfactant QRMAE Can Be Utilized as an Amorphous Aggregate Inducer: A Case Study of Mammalian Serum Albumin

Quaternary amine of diethylaminoethyl rosin ester (QRMAE), chemically synthesized biocompatible rosin based cationic surfactant, has various biological applications including its use as a food product additive. In this study, we examined the amorphous aggregation behavior of mammalian serum albumins at pH 7.5, i.e., two units above their isoelectric points (pI ~5.5), and the roles played by positive charge and hydrophobicity of exogenously added rosin surfactant QRMAE. The study was carried out on five mammalian serum albumins, using various spectroscopic methods, dye binding assay, circular dichroism and electron microscopy. The thermodynamics of the binding of mammalian serum albumins to cationic rosin modified surfactant were established using isothermal titration calorimetry (ITC). It was observed that a suitable molar ratio of protein to QRMAE surfactant enthusiastically induces amorphous aggregate formation at a pH above two units of pI. Rosin surfactant QRMAE-albumins interactions revealed a unique interplay between the initial electrostatic and the subsequent hydrophobic interactions that play an important role towards the formation of hydrophobic interactions-driven amorphous aggregate. Amorphous aggregation of proteins is associated with varying diseases, from the formation of protein wine haze to the expansion of the eye lenses in cataract, during the expression and purification of recombinant proteins. This study can be used for the design of novel biomolecules or drugs with the ability to neutralize factor(s) responsible for the aggregate formation, in addition to various other industrial applications.