分子印迹技术在多肽、蛋白质分离中的应用

介绍了分子印迹聚合物(Molecularly Imprinted Ploymer,MIP)在分离生物大分子——多肽、蛋白质等领域中的应用进展，及其发展前景。     

草鱼日粮中虾蛋白肽对幼龄草鱼生长性能的影响

用300尾(3.74±0.23)g的草鱼Ctenopharyngodon idella作日粮中不同添加量虾蛋白肽(平均肽链长为3.61)对幼龄草鱼生长性能影响研究.实验分为4组,每组75尾鱼,每25尾鱼饲于0.25m3的水族箱中,每组设3个平行水族箱.试验1、2、3组日粮中分别添加w为0.25%虾蛋白肽 w为0.75%鱼粉、w为0.5%虾蛋白肽 w为0.5%鱼粉、w为1%虾蛋白肽,对照组添加w为1%鱼粉.试验期56 d.结果表明:试验各组草鱼其特定生长率、蛋白保留效率、饲料系数和血浆中磷、镁含量与小肽总量均高于对照组,特别是试验2、3组与对照组有显著性差异(P＜O.05);草鱼日粮中适宜的虾蛋白肽添加量为w为0.5%.草鱼日粮中添加一定比例的虾蛋白肽均可提高饲料表观消化率和蛋白消化率,增加血液循环中生物活性肽的含量,提高机体对日粮中氨基酸的利用率,从而增加体内氮沉积,减少肝胰脏和肠系膜脂肪储积.     

Proteolytic generation and aggregation of peptides from transmembrane regions: lung surfactant protein C and amyloid β-peptide

The formation of amyloid fibrils is associated with several devastating diseases in humans and animals, including e.g. Alzheimers disease (AD) and the spongiform encephalopathies. Here, we review and discuss the current knowledge on two amyloid peptides: lung surfactant protein C (SP-C) and the amyloid -peptide (A), implicated in human lung disease and in AD, respectively. Both these hydrophobic peptides are derived from the transmembrane region of their precursor protein, and can transit from a monomeric -helical state to a -sheet fibril. The helices of SP-C and A are composed of amino acid residues with inherently higher propensities for strand than helix conformation. Their helical states are stabilized by a membrane environment, and loss of membrane association thus promotes structural conversion and fibril formation. We speculate that the loss of structural context for sequences with a high propensity for formation of sheets may be a common feature of amyloid formation in general.Received 9 July 2003; received after revision 15 August 2003; accepted 3 September 2003     

Molecular modeling of the ion channel-like nanotube structure of amyloid β-peptide

The ion channel-like nanotube structure of the oligomers of amyloid β-peptide (Aβ) was first investi-gated by molecular modeling. The results reveal that the hydrogen bond net is one of the key factors to stabilize the structure. The hydrophobicity distribution mode of the side chains is in favor of the structure inserting into the bilayers and forming a hydrophilic pore. The lumen space is under the control of the negative potential,weaker but spreading continuously,to which the cation selectivity attributes; meanwhile,the alternate distribution of the stronger positive and negative potentials makes the electrostatic distribution of the structure framework balance,which is also one of the key factors stabilizing the structure. The results lay the theoretical foundation for illuminating the structure stability and the ion permeability,and give a clue to elucidating the molecular mechanism of Alzheimer's dis-ease (AD) and designing novel drugs to prevent or reverse AD at the root.     

复合酶法水解花生粕制备抗氧化肽的工艺优化

以DPPH自由基清除率和收率为响应值,采用响应面法对复合酶法水解花生粕生产抗氧化性肽工艺条件进行了优化.研究结果表明,碱性蛋白酶和中性蛋白酶对花生粕具有较强的水解能力,二者以2∶1的比例对花生粕水解时,较优水解条件是50℃,pH值8.54,底物质量分数8.34%.在此条件下酶解16 h,花生肽收率为65.80%,在花生肽质量浓度为0.55 mg/mL时,对DPPH自由基清除率为25.77%,与优化前相比,复合酶比碱性蛋白酶单酶水解时收率和DPPH自由基清除率分别提高了7.48%和7.66%,比中性蛋白酶单酶水解时分别提高了10.29%和22.53%.     

原小檗碱类多靶点抗老年性痴呆症药理作用

 原小檗碱是一类异喹啉生物碱,具有相同的药效基团。本研究检测了以小檗碱和四氢黄藤素为代表的原小檗碱多靶点抗AD效应及分子作用机制。以人神经母细胞瘤SH SY5Y为研究对象,利用Western blot检测药物对细胞全长APP和BACE1蛋白水平的影响;ELISA技术检测细胞外可溶性淀粉样前体蛋白α (sAPPα) 的分泌;酶活性检测试剂盒检测乙酰胆碱酯酶 (AChE) 的活性。结果显示,药物处理细胞24 h后,二者均能显著地抑制BACE1蛋白的表达,并可使sAPPα的分泌显著增加,但未见细胞APP蛋白含量明显变化,同时对该细胞的AchE的活性具有显著的抑制作用。以上结果证明：原小檗碱生物碱药理活性涉及预防和减少Aβ沉积和增强乙酰胆碱对脑胆碱受体的作用,并通过细胞自身的代谢增加具有神经营养作用的sAPPα的分泌。提示此类化合物具有多靶点抗老年痴呆的潜能,通过结构优化改造,将有望获得副作用小的新一代抗AD有效药物。     

Tauroursodeoxycholic acid prevents E22Q Alzheimer’s Aβ toxicity in human cerebral endothelial cells

The vasculotropic E22Q mutant of the amyloid-β (Aβ) peptide is associated with hereditary cerebral hemorrhage with amyloidosis Dutch type. The cellular mechanism(s) of toxicity and nature of the AβE22Q toxic assemblies are not completely understood. Comparative assessment of structural parameters and cell death mechanisms elicited in primary human cerebral endothelial cells by AβE22Q and wild-type Aβ revealed that only AβE22Q triggered the Bax mitochondrial pathway of apoptosis. AβE22Q neither matched the fast oligomerization kinetics of Aβ42 nor reached its predominant β-sheet structure, achieving a modest degree of oligomerization with a secondary structure that remained a mixture of β and random conformations. The endogenous molecule tauroursodeoxycholic acid (TUDCA) was a strong modulator of AβE22Q-triggered apoptosis but did not significantly change the secondary structures and fibrillogenic propensities of Aβ peptides. These data dissociate the pro-apoptotic properties of Aβ peptides from their distinct mechanisms of aggregation/fibrillization in vitro, providing new perspectives for modulation of amyloid toxicity. Received 20 November 2008; received after revision 12 December 2008; accepted 12 January 2009     

We find them here, we find them there: Functional bacterial amyloid

Protein amyloid is often deposited in connection with neurodegenerative diseases. Such deposits generally possess three principal drawbacks: cytotoxicity, lack of spatial control in their deposition and structural polymorphism. These are typical features of biologically non-optimized systems which have not been exposed to evolutionary pressure. Nevertheless, Nature uses the cross-beta self-organizing principle in many structural contexts where a strong but pliable material is needed. Functional amyloid is found in humans, invertebrates, fungi and, not least, bacteria, in which amyloid may be the rule rather than the exception. Detailed case studies reveal how directed nucleation can use tailor-made proteins optimized to assume a specific amyloid conformation, leading to remarkably robust assemblies. This makes it highly challenging to purify and analyze the products formed in vivo. We contrast pathogenic and in-vitro-formed amyloid with functional amyloid, paying particular reference to bacterial amyloid, and discuss challenges and perspectives in identifying and characterizing this class of protein.     

降钙素基因相关肽和内皮素-1对支气管上皮细胞与嗜酸性粒细胞黏附功能的影响

观察肺内调节肽对嗜酸性粒细胞(EOS)与支气管上皮细胞(BEC)粘附功能的影响.选择降钙素基因相关肽(CGRP)和内皮素-1(ET-1)两种调节肽作为研究对象,观察它们对BEC白细胞介素(ILs)分泌及细胞间粘附分子-1(ICAM-1)表达的影响.结果表明:CGRP,ET-1可使O3应激的BEC分泌ILs能力升高,并上调气道上皮ICAM-1的表达.CGRP、ET-1能促进BEC与EOS的粘附,对气道炎症起正性调节作用,加重局部的损伤.