红小豆叶片硝酸还原酶测定法的改进

以红小豆资源为材料,采用硝酸还原酶测定试剂盒测定各材料硝酸还原酶含量。实验结果发现:严格按照试剂盒说明操作,普遍存在对照管值与测定管值相差不大,或对照管值大于测定管值的非正常现象。经过诱导硝酸还原酶时间梯度实验、37℃保温时间梯度实验,否认了此现象产生是由酶活性表达过低引起。将对照管中成分进行调整,消除了该现象。经讨论,认为调整后的对照比原对照更为合理。     

钼对葡萄叶片硝酸还原酶活性及生长发育的影响

适量施钼有利于葡萄生长过程中硝酸还原酶活性的提高，缺乏或过量则使其降低，其他微量元素对ＮＲ活性也产生一定影响。由于酶活性的差异也对葡萄的生长发育带来影响。     

渗透胁迫对小麦幼苗硝酸还原酶活性的影响

本文研究了小麦幼苗根部和离体叶切段遭受PEG(MW=6000)渗透胁迫后叶内硝酸还原酶(NR)活性的变化,发现渗透胁迫不仅降低叶片中已有的NR活性,还抑制底物NO_3~-对NR的诱导过程。进一步研究表明,幼苗NR的诱导受抑与渗透胁迫下叶内诱导物不足有关。     

Subatomic and atomic crystallographic studies of aldose reductase: implications for inhibitor binding

The determination of several of aldose reductase-inhibitor complexes at subatomic resolution has revealed new structural details, including the specific interatomic contacts involved in inhibitor binding. In this article, we review the structures of the complexes of ALR2 with IDD 594 (resolution: 0.66 Å, IC50 (concentration of the inhibitor that produced half-maximal effect): 30 nM, space group: P2¹), IDD 393 (resolution: 0.90 Å, IC50: 6 nM, space group: P1), fidarestat (resolution: 0.92 Å, IC50: 9 nM, space group: P2¹) and minalrestat (resolution: 1.10 Å, IC50: 73 nM, space group: P1). The structures are compared and found to be highly reproductible within the same space group (root mean square (RMS) deviations: 0.15 0.3 Å). The mode of binding of the carboxylate inhibitors IDD 594 and IDD 393 is analysed. The binding of the carboxylate head can be accurately determined by the subatomic resolution structures, since both the protonation states and the positions of the atoms are very precisely known. The differences appear in the binding in the specificity pocket. The high-resolution structures explain the differences in IC50, which are confirmed both experimentally by mass spectrometry measures of VC50 and theoretically by free energy perturbation calculations. The binding of the cyclic imide inhibitors fidarestat and minalrestat is also described, focusing on the observation of a Cl^- ion which binds simultaneously with fidarestat. The presence of this anion, binding also to the active site residue His110, leads to a mechanism in which the inhibitor can bind in a neutral state and then become charged inside the active site pocket. This mechanism can explain the excellent in vivo properties of cyclic imide inhibitors. In summary, the complete and detailed information supplied by the subatomic resolution structures can explain the differences in binding energy of the different inhibitors.     

突变型大肠杆菌二氢叶酸还原酶基因的合成

利用PCR定点突变技术，以野生型大肠杆菌二氢叶酸还原酶基因为模板，获取3种突变型(Cys85Ser．Cysl51Ser，Cys85／151Ser)二氢叶酸还原酶(DHFR)基因．用限制性内切酶BamH Ⅰ与Pst Ⅰ将3种突变基因片段插入到克隆载体pUC18上，进行蓝白筛选，将筛选的克隆进行DNA序列测定．     

脱落酸对小麦幼苗各器官中硝酸还原酶活力的影响

在含有４μＭ脱落酸（ＡＢＡ）的Ｈｏａｇｌａｎｄ培养液中培养的小麦幼苗，在离休条件下，根、茎、叶中的硝酸还原酶（ＮＲ），＊力均受到抑制，在处理２１～４８ｈ期间，抑制达到高峰，７２ｈ根、叶中的活性得到恢复，９６ｈ茎中的活性恢复。除去培养液中的ＡＢＡ２４ｈ后，继续观察实验的结果表明，ＡＢＡ对ＮＲ活力的抑制不依赖于硝酸盐的吸收，尽管ＡＢＡ降低了叶片中硝酸盐的含量，但在组织的硝酸盐水平与ＡＢＡ之间没有直接的关系．     

渗透胁迫等因素对春小麦幼苗硝酸还原酶活性的影响

本文研究了渗透胁迫、热胁迫、外源脯氨酸和脱落酸对春小麦幼苗 NR 活性的影响.结果表明,NR 对水分胁迫极为敏感,随胁强增加,活性持续下降;解除胁迫后,一般24h 内活性即能恢复.NR 对热胁迫也很敏感,当温度高于正常生长温度10℃时,活性即迅速下降。外源脯氨酸能够减缓胁迫条件下 NR 活性的下降。外源脱落酸在胁迫条件下加剧 NR 活性的下降。     

黄米醇溶蛋白对小鼠胆固醇代谢的调节作用

比较了分别饲喂酪蛋白、大豆分离蛋白、黄米醇溶蛋白小鼠的血清总胆固醇、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇,甘油三酯浓度和动脉硬化指数,旨在评价黄米醇溶蛋白对小鼠胆固醇代谢的特定作用。结果显示:黄米醇溶蛋白(添加1.82%Lys和0.23%Trp)能显著提高小鼠血清高密度脂蛋白胆固醇浓度(P<0.05),降低动脉硬化指数;但体外实验表明:黄米醇溶蛋白不能抑制羟甲基戊二酸单酰辅酶A(HMG-CoA)还原酶的活性,即不能抑制胆固醇的合成。线性回归分析得出黄米醇溶蛋白对小鼠血清胆固醇代谢的调节作用与其蛋氧酸与甘氨酸含量之比、疏水性氨基酸、含硫氨基酸含量密切相关。     

硫氧还蛋白还原酶结构分形研究

提出并应用改进的分形方法对硫氧还蛋白还原酶(TrxR)结构进行研究。建立了适用于TrxR的分形及数据处理方法。发现正常的TrxR的结构分维值约为1.33,氧化后结构分维值增大。提出了蛋白质结构分维值是表征蛋白质分子状态的重要特征之一的观点,并结合TrxR验证了该观点。提出了药物分子分维值应与靶酶的结构分维值相契合的观点,并结合以TrxR为靶点的药物研发实验验证了该观点。