Zinc binding to peptide analogs of the structural zinc site in alcohol dehydrogenase: Implications for an entatic state

Zinc binding to the peptide replica and analogs to residues 93–115 of horse liver alcohol dehydrogenase (ADH) was examined by competition of the peptides and the chromophoric chelator 4-(2- pyridylazo)resorcinol for zinc and X-ray absorption fine structure analysis of the zinc ligands. In the enzyme, zinc is coordinated by four Cys residues. In the peptide replica, zinc is bound to three Cys and one His residue. A four-Cys zinc coordination is observed only when His is removed, leading to increased zinc stability. ADH crystal structures reveal that the ε-amino group of the conserved residue Lys323 is within H-bond distance of the backbone amide oxygens of residues 103, 105 and 108, likely stabilizing the zinc coordination in the enzyme. The peptide data thus indicate structural strain and increased energy in the zinc-binding site in the protein, characteristic of an entatic state, implying a functional nature for this zinc site. Received 3 July 2008; received after revision 11 August 2008; accepted 1 September 2008     

外源性H2S通过调节β-位淀粉样前体蛋白裂解酶1表达对PC12细胞APP／Aβ代谢的影响

目的观察外源性硫化氢对嗜铬细胞瘤细胞β-位淀粉样前体蛋白裂解酶1（BACE1）的调节作用,进而探讨其对淀粉样前体蛋白／β-位淀粉样蛋白代谢途径的影响。方法用硫氢化钠作外源性H2s供体,实验设空白对照组、NaHs50μmol／L组、NaHS100μmol／L组和NariS200μmol／L组,按分组浓度处理PC12细胞24h后,RT-PCR和Western blot法检测细胞内BACE1 mRNA及蛋白表达,并用Western blot法继而检测APP代谢过程中关键蛋白APP、C99、C83表达变化,EusA法检测细胞培养液中AB40和AB42水平。结果NaHS在实验浓度范围内从基因与蛋白两个水平上呈剂量依赖性下调BACE1表达,并下调C99、Ap40和Ap42蛋白表达,上调C83蛋白,各NaHS组分别与对照组比较,差别均有统计学意义（P〈0．05）,而对APP蛋白表达没有影响,各组间比较差别无显著性（P〉0．05）。结论外源性H2s具有通过调节PC12细胞BACE1表达下调APP／Aβ代谢的作用。     

Transthyretin: a review from a structural perspective

Transthyretin (formerly called prealbumin) plays important physiological roles as a transporter of thyroxine and retinol-binding protein. X-ray structural studies have provided information on the active conformation of the protein and the site of binding of both ligands. Transthyretin is also one of the precursor proteins commonly found in amyloid deposits. Both wild-type and single-amino-acid-substituted variants have been identified in amyloid deposits, the variants being more amyloidogenic. Sequencing of the gene and the resulting production of a transgenic mouse model have resulted in progress toward solving the mechanism of amyloid formation and detecting the variant gene in individuals at risk. Received 23 January 2001; received after revision 4 April 2001; accepted 30 April 2001     

Presence of hepcidin-25 in biological fluids: Bile,ascitic and pleural fluids

AIM: To examine body fluids such as ascitic fluid (AF),saliva,bile and pleural effusions for the presence of hepcidin using a novel radioimmunoassay (RIA).METHODS: Serum samples were collected from 25 healthy volunteers (mean age: 36 ± 11.9 years,11 males,14 females).In addition bile was obtained from 12 patients undergoing endoscopic retrograde cholangiopancreatography (mean age: 66.9 ± 16.7 years,M:F = 5:7).Saliva was collected from 17 healthy volunteers (mean age: 35 ± 9.9 years,M:F = 8:9).Pleural and AF...     

Application of surface plasmon resonance in screening adsorbents and explaining adsorption phenomena using model polymers

A surface plasmon resonance (SPR) technique was developed as an online screening method to identify effective adsorbents for the peptide NH₂-Cys-Thr-Trp-Trp-COOH (CW-4). To validate the SPR technique, several linear polymers containing amino acid residues were synthesized as models of their corresponding adsorbents. SPR screening of all the linear polymers demonstrated that the linear polymer containing phenylalanine residues (PPhe) exhibited the highest affinity for CW-4 among the polymers tested. In accordance with the screening results from SPR, the adsorbent containing phenylalanine (APhe) exhibited the highest adsorption capacity for CW-4. An interesting observation was that the amounts of CW-4 adsorbed on 3 adsorbents were quite small, even though both SPR and isothermal titration calorimetry (ITC) experiments indicated the presence of interactions between CW-4 and the three polymers. A kinetic analysis performed using SPR technology suggested such a phenomenon was ascribed to the poor stability of the CW-4/polymer complexes; that is, CW-4 associates with the polymers quickly, but it also dissociated quickly. The combined results suggested that SPR was a promising tool to identify the optimal adsorbent for peptides, analyze the interactions contributing to adsorption and explain adsorption phenomena using polymer models.     

The effect of corticotropin-releasing factor and pro-opiomelanocortin-derived peptides on the phagocytosis of molluscan hemocytes

The effect of corticotropin-releasing factor (CRF) and pro-opiomelanocortin (POMC)-derived peptides on hemocyte phagocytosis in two molluscs,Planorbarius corneus andViviparus ater was studied. The peptides and related fragments examined are those which have been shown to influence hemocyte motility in the two species. The results obtained revealed that the effects on phagocytosis are not directly correlated with previous findings on cell motility. Furthermore, the mode of action of an individual peptide could be species-specific and dose-dependent. The relationships between peptides, locomotion and phagocytosis in these molluscs are discussed.     

Molecular pathogenesis of apolipoprotein E-mediated amyloidosis in late-onset Alzheimer’s disease

Apolipoprotein E (apoE) ɛ4 allele is a genetic risk factor for late-onset familial and sporadic Alzheimer’s disease (AD). In the central nervous system, apoE is secreted mainly by astrocytes as a constituent of high-density lipoproteins. A recent study using apoE knockout mice provided strong evidence that apoE promotes cerebral deposition of amyloid β protein (Aβ). However, no clear explanation of the pathogenesis of apoE-induced AD has been provided. Here we discuss two possible mechanisms by which apoE might enhance Aβ deposition. One is the intracellular pathway in which apoE is internalized by neurons and induces lysosomal accumulation of Aβ and amyloidogenic APP (amyloid precursor protein) fragments, leading to neuronal death. The other is the extracellular pathway in which apoE-containing lipoproteins are trapped by Aβ1–42 deposits mobilizing soluble Aβ peptides and consequently enlarge amyloid plaques. These two mechanisms may operate at different stages of AD pathogenesis and suggest a chaperone-like function for the apoE molecule. Received 4 February 1999; received after revision 9 April 1999; accepted 23 April 1999     

三种脑-肠肽对人绒毛膜促性腺激素分泌的影响

本工作利用灌流技术观察了3种脑-肠肽(P物质,CCK及NT)对新鲜人工流产的绒毛hCG分泌的影响。结果表明,P物质,CCK及NT均明显刺激绒毛释放hCG,并发现CCK作用大于P物质及NT。提示,hCG的分泌可能受这3种脑-肠肽的调节。     

抗菌肽的抗菌机制及其在医药中的应用前景

随着抗生素副作用的日益凸显,以及其被批准数量的逐年下降,研发功能相似且不易产生耐药性的其他活性物质迫在眉睫。抗菌肽具有广谱抗菌性、热稳定性,作用机理独特,既能起到体内抗病菌感染、促进癌细胞凋亡的作用,还可作为抗病毒化疗剂和医疗器械的抗菌材料。系统综述了抗菌肽的抗菌机制,及其在医药领域抑制抗耐药菌的产生、抗癌以及抗病毒感染的作用,阐明了抗菌肽存在的稳定性、溶血性及毒性等问题,并对其发展前景进行了展望。指出今后可在以下方面对抗菌肽展开深入研究:1)结合体内抑菌机制进行研究,推动抗菌肽的开发应用;2)通过替换、修饰氨基酸残基,合成杂合肽,优化抗菌肽结构,提高其活性,降低毒副作用并预防耐药性的产生;3)利用抗菌肽传递系统或抗菌肽与绿色天然提取物联合作用,提高抗菌活性和生物利用度。