室温下N-羟基琥珀酰亚胺活化酯法合成吡咯-2,5-二乙酰胺

在室温下用一锅反应法以二环己基碳二亚胺（DCC）（2 equiv．）为偶合试剂，使用过量（4equiv．）的胺，用N-羟基琥珀酰亚胺（Hosu）（2 equiv．）活化酯法合成了吡咯-2，5-二酰胺，且产率（70％）比在低温时（63％）有所提高．探讨了选用不同的合成方法及偶合试剂以及反应条件和试剂的用量对合成目标产物的影响．     

异柠檬酸裂解酶肽类抑制剂结构的修饰

利用Fmoc固相多肽合成法, 按已知异柠檬酸裂解酶抑制剂的直链肽氨基酸序列合成首尾相连的环肽抑制剂. 经色谱纯化和质谱鉴定, 其相对分子质量的实测值与理论值相符. 抑制率实验结果表明, 合成的环肽对异柠檬酸裂解酶有明显抑制作用, 抑制率大于50%. 采用高效液相色谱法分别检测直链肽和环肽在血浆中的半衰期, 实验
结果表明, 环肽在血浆中的半衰期为11 min, 比直链肽的半衰期延长175%.     

从七肽噬菌体展示库中筛选蛋白质配基

为了研究噬菌体展示技术的应用，以溶菌酶为靶分子从七肽噬菌体展示库中筛选蛋白质的高亲和力噬菌体配体，所筛选的亲和力最高的噬菌体的ELISA检测值A405nm可达0．634．通过比较亲和性噬菌体外源插入肽的DNA序列，认为基元HWWW是肽段与酶分子发生亲和的必需序列．此外，由于靶分子和高亲和性展示肽的等电点分别为11．2和6．74，因此在亲和环境中携带异种电荷，利于亲和吸附的发生，而此时低亲和性展示肽与靶分子携带同种电荷，阻碍了亲和吸附．同时，高亲和性肽段HWWPAS和与其有较高同源性的肽段HWTWWNL都有适中的疏水性，这有利于肽与靶分子表面的疏水位点相互作用从而产生亲和吸附．     

鱼类尾部神经分泌系统研究进展

比较详细地介绍了自１９世纪末到本世纪末近一个世纪生物学家们在鱼类尾部神经分泌系统的形态结构及组成,其所分泌的活性肽及生理效应,该系统的生理功能和分泌方式及神经支配等方面的研究成果和发展趋势,为全面了解该系统的结构和功能提供了较为翔实的资料,并为进一步深入研究该系统了方向和方法。     

Evaluation of TILI-2 as an Anti-Tyrosinase,Anti-Oxidative Agent and Its Role in Preventing Melanogenesis Using a Proteomics Approach

There is a desire to develop new molecules that can combat hyperpigmentation. To this end, the N-terminal cysteine-containing heptapeptide TILI-2 has shown promising preliminary results. In this work, the mechanism by which it works was evaluated using a series of biochemical assays focusing on known biochemical pathways, followed by LC-MS/MS proteomics to discover pathways that have not been considered before. We demonstrate that TILI-2 is a competitive inhibitor of tyrosinase’s monophenolase activity and it could potentially scavenge ABTS and DPPH radicals. It has a very low cytotoxicity up to 1400 µM against human fibroblast NFDH cells and macrophage-like RAW 264.7 cells. Our proteomics study revealed that another putative mechanism by which TILI-2 may reduce melanin production involves the disruption of the TGF-β signaling pathway in mouse B16F1 cells. This result suggests that TILI-2 has potential scope to be used as a depigmenting agent.     

The Effects of Charged Amino Acid Side-Chain Length on Diagonal Cross-Strand Interactions between Carboxylate- and Ammonium-Containing Residues in a β-Hairpin

The β-sheet is one of the common protein secondary structures, and the aberrant aggregation of β-sheets is implicated in various neurodegenerative diseases. Cross-strand interactions are an important determinant of β-sheet stability. Accordingly, both diagonal and lateral cross-strand interactions have been studied. Surprisingly, diagonal cross-strand ion-pairing interactions have yet to be investigated. Herein, we present a systematic study on the effects of charged amino acid side-chain length on a diagonal ion-pairing interaction between carboxylate- and ammonium-containing residues in a β-hairpin. To this end, 2D-NMR was used to investigate the conformation of the peptides. The fraction folded population and the folding free energy were derived from the chemical shift data. The fraction folded population for these peptides with potential diagonal ion pairs was mostly lower compared to the corresponding peptide with a potential lateral ion pair. The diagonal ion-pairing interaction energy was derived using double mutant cycle analysis. The Asp2-Dab9 (Asp: one methylene; Dab: two methylenes) interaction was the most stabilizing (−0.79 ± 0.14 kcal/mol), most likely representing an optimal balance between the entropic penalty to enable the ion-pairing interaction and the number of side-chain conformations that can accommodate the interaction. These results should be useful for designing β-sheet containing molecular entities for various applications.     

高效液相色谱/质谱法识别不同明胶酶解产物中特征多肽

在不同动物胶原蛋白序列比对基础上,利用高效液相色谱/质谱联用技术(HPLC/MS)建立一种通过特征多肽进行明胶鉴别的方法。实验以牛明胶和猪明胶为对象,序列比对结果表明,牛和猪的Ⅰ型胶原蛋白α1链和α2链均存在氨基酸序列差异。利用胰蛋白酶将牛明胶和猪明胶降解,通过HPLC/MS分析了两种明胶酶解产物中的多肽片段。结果表明:牛明胶和猪明胶酶解产物中均可检测出存在序列差异的特征性多肽,其氨基酸序列差异与两种胶原蛋白序列比对结果中的序列差异一致;酶解产物中特征多肽上的脯氨酸存在不同程度的羟基化修饰,特征多肽的相对含量主要受明胶分子量范围影响。利用HPLC/MS检测明胶酶解产物中的特征多肽是一种鉴别牛明胶和猪明胶的有效方法。     

修饰合成冬鲽抗冻肽基因的设计和克隆

动物界和植物界密码子使用频率不同,冬Ｄｉｅ抗冻肽中丙氨酸占６０％而且偏爱ＧＣＣ,３８个Ａｌａ密友子中２３个为ＧＣＣ。我们设计合成抗冻肽基因时采用了植物基因常用密码子,用甘薯信号肽序列取了抗冻肽的ｐｒｅ序列,省略了Ｐｒｏ序列,并 信号肽下游二、四、八拷贝正向串联的成太的修饰基因,经测序证实与设计完成一致。利用ＱＩＡ表达系统,在大肠杆菌中实现了ＤＨＦＲ－成熟抗冻肽单体和二体的融合表达。     

通过pR1aS导肽提高植物中绿色荧光蛋白的激发强度

绿色荧光蛋白在植物细胞胞质表达中对转化植物细胞的再生存在不利的影响,为增强绿色荧光蛋白在植物细胞中的表达,在mgfp的5'端连接了pR1aS信号肽序列,同时在3'末端引入滞留在内质网中的特异序列KDEL,成功构建了植物表达载体pBLG-pR1aS-gfp,经转基因烟草表达分析,结果表明:转化体植株显著增加了绿色荧光蛋白在烟草中的表达,同时消除了绿色荧光蛋白对植物潜在的光毒害。这为植物转基因转化频率的研究和转基因植物安全性评价提供了一种有利的工具。     

姬松茸菌丝体中活性肽提取工艺的初步研究

通过对姬松茸菌丝体中活性肽提取工艺的初步研究，并采用单因素考察及正交实验法对姬松茸茵丝体的活性肽提取条件进行了优化。结果：在姬松茸干菌丝粉中加入8倍体积的水，在温度28℃和pH9．0的条件下，用70％乙醇的沉淀提取6h，所得提取液中活性肽的含量及抗氧化性均最优，经毛细管电泳图谱分析其活性肽分布也较适宜。