Constraining an Irregular Peptide Secondary Structure through Ring‐Closing Alkyne Metathesis

Macrocyclization can be used to constrain peptides in their bioactive conformations, thereby supporting target affinity and bioactivity. In particular, for the targeting of challenging protein–protein interactions, macrocyclic peptides have proven to be very useful. Available approaches focus on the stabilization of α‐helices, which limits their general applicability. Here we report for the first time on the use of ring‐closing alkyne metathesis for the stabilization of an irregular peptide secondary structure. A small library of alkyne‐crosslinked peptides provided a number of derivatives with improved target affinity relative to the linear parent peptide. In addition, we report the crystal structure of the highest‐affinity derivative in a complex with its protein target 14‐3‐3ζ. It can be expected that the alkyne‐based macrocyclization of irregular binding epitopes should give rise to new scaffolds suitable for targeting of currently intractable proteins.     

X-ray Crystallographic Structure of α-Helical Peptide Stabilized by Hydrocarbon Stapling at i,i + 1 Positions

Hydrocarbon stapling is a useful tool for stabilizing the secondary structure of peptides. Among several methods, hydrocarbon stapling at i,i + 1 positions was not extensively studied, and their secondary structures are not clarified. In this study, we investigate i,i + 1 hydrocarbon stapling between cis-4-allyloxy-l-proline and various olefin-tethered amino acids. Depending on the ring size of the stapled side chains and structure of the olefin-tethered amino acids, E- or Z-selectivities were observed during the ring-closing metathesis reaction (E/Z was up to 8.5:1 for 17–14-membered rings and up to 1:20 for 13-membered rings). We performed X-ray crystallographic analysis of hydrocarbon stapled peptide at i,i + 1 positions. The X-ray crystallographic structure suggested that the i,i + 1 staple stabilizes the peptide secondary structure to the right-handed α-helix. These findings are especially important for short oligopeptides because the employed stapling method uses two minimal amino acid residues adjacent to each other.     

Aphid BCR4 Structure and Activity Uncover a New Defensin Peptide Superfamily

Aphids (Hemiptera: Aphidoidea) are among the most detrimental insects for agricultural plants, and their management is a great challenge in agronomical research. A new class of proteins, called Bacteriocyte-specific Cysteine-Rich (BCR) peptides, provides an alternative to chemical insecticides for pest control. BCRs were initially identified in the pea aphid Acyrthosiphon pisum. They are small disulfide bond-rich proteins expressed exclusively in aphid bacteriocytes, the insect cells that host intracellular symbiotic bacteria. Here, we show that one of the A. pisum BCRs, BCR4, displays prominent insecticidal activity against the pea aphid, impairing insect survival and nymphal growth, providing evidence for its potential use as a new biopesticide. Our comparative genomics and phylogenetic analyses indicate that BCRs are restricted to the aphid lineage. The 3D structure of BCR4 reveals that this peptide belongs to an as-yet-unknown structural class of peptides and defines a new superfamily of defensins.     

A Conditionally Fluorescent Peptide Reporter of Secondary Structure Modulation

Proteins containing intrinsic disorder often form secondary structure upon interaction with a binding partner. Modulating such structures presents an approach for manipulating the resultant functional outcomes. Translational repressor protein 4E-BP1 is an example of an intrinsically disordered protein that forms an α-helix upon binding to its protein ligand, eIF4E. Current biophysical methods for analyzing binding-induced structural changes are low-throughput, require large amounts of sample, or are extremely sensitive to signal interference by the ligand itself. Herein, we describe the discovery and development of a conditionally fluorescent 4E-BP1 peptide that reports structural changes of its helix in high-throughput format. This reporter peptide is based on conditional quenching of fluorescein by thioamides. In this case, fluorescence signal increases as the peptide becomes more ordered. Conversely, destabilization of the α-helix results in decreased fluorescence signal. The low concentration and low volume of peptide required make this approach amenable for high-throughput screening to discover ligands that alter peptide secondary structure.     

微波辐射在多肽合成中的研究进展

微波作为一种可以加速有机反应的辅助技术已广泛地应用于多肽及其衍生物的合成。简要介绍了微波辐射加速有机反应的机理,综述了微波辐射在多肽及其衍生物合成中的最新研究进展,并展望了微波在多肽合成中的前景。     

Biosynthesis of the Peptide Antibiotic Feglymycin by a Linear Nonribosomal Peptide Synthetase Mechanism

Feglymycin, a peptide antibiotic produced by Streptomyces sp. DSM 11171, consists mostly of nonproteinogenic phenylglycine‐type amino acids. It possesses antibacterial activity against methicillin‐resistant Staphylococcus aureus strains and antiviral activity against HIV. Inhibition of the early steps of bacterial peptidoglycan synthesis indicated a mode of action different from those of other peptide antibiotics. Here we describe the identification and assignment of the feglymycin (feg) biosynthesis gene cluster, which codes for a 13‐module nonribosomal peptide synthetase (NRPS) system. Inactivation of an NRPS gene and supplementation of a hydroxymandelate oxidase mutant with the amino acid l ‐Hpg proved the identity of the feg cluster. Feeding of Hpg‐related unnatural amino acids was not successful. This characterization of the feg cluster is an important step to understanding the biosynthesis of this potent antibacterial peptide.     

受阻胺光稳定剂的多功能性及其结构优化

简述了在高分子材料中受阻胺光稳定剂（HALS）作用的机理以及其他功能,并介绍了目前从结构上对受阻胺光稳定剂进行的性能优化的4种主要途径：高相对分子质量化,引入其他功能基团,低碱性化和提高相容性。     

片段法合成多肽的研究

以片段合成法和直接合成法分别合成一个比较长的多肽,并用质谱和HPLC进行检测,结果显示用片段合成法合成的多肽,其合成效率、合成纯度均明显高于直接法。     

十六烷基三甲基溴化铵与亮绿SF作用的共振光散射研究

研究了十六烷基三甲基溴化铵(CTAB)和亮绿SF作用的共振散射光谱特征。结果表明:在p H=4.10缓冲介质中,十六烷基三甲基溴化铵与亮绿SF相互作用形成离子缔合物,产生以505 nm为特征峰的共振光散射(RLS)增强信号,其RLS增强程度与亮绿SF浓度成线性关系,检出限和线性范围分别为97.6 nmol/L和0.98~75μmol/L,据此建立了痕量亮绿SF的共振光散射分析方法。将方法用于水样中亮绿SF含量的快速检测,结果令人满意。     

Selective Covalent Targeting of Anti-apoptotic BFL-1 by a Sulfonium-Tethered Peptide

Anti-apoptotic B cell lymphoma 2 (BCL-2) family proteins are proven targets for human cancers. Targeting the BH3-binding pockets of these anti-apoptotic proteins could reactivate apoptosis in BCL-2-depedent cancers. BFL-1 is a BCL-2 family protein overexpressed in various chemoresistant cancers. A unique cysteine at the binding interface of the BH3 and BFL-1 was previously proven to be an intriguing targeting site to irreversibly inhibit BFL-1 functions with stabilized cyclic peptide bearing a covalent warhead. Recently, we developed a sulfonium-tethered peptide cyclization strategy to construct peptide ligands that could selectively and efficiently react with the cysteine(s) of target proteins near the interacting interface. Using this method, we constructed a BFL-1 peptide inhibitor, B4-MC, that could selectively conjugate with BFL-1 both in vitro and in cell. B4-MC showed good cellular uptake, colocalized with BFL-1 on mitochondria, and showed obvious growth inhibition of BFL-1 over-expressed cancer cell lines.     

DBC-偶氮羧共振光散射法测定蛋白质

基于蛋白质对DBC-偶氮羧光散射增强的效应,拟定了一种测定蛋白质的新方法.在pH 4.1的Britton -Robinson(B-R)缓冲溶液中,蛋白质与DBC-偶氮羧结合,产生强烈的共振光散射.在362 nm处,共振光散射强度较大,且光散射强度与加入的蛋白质浓度在一定范围内呈线性关系,其中牛血清白蛋白(BSA)在0.05～7 mg·L-1、人血清白蛋白(HSA)在0.05～8 mg·L-1、溶菌酶(Lyso) 在0.05～7 mg·L-1.该法用于人血清总蛋白含量的测定,结果令人满意.     

紫外光对桑蚕丝结构及性能影响研究

文章以普通商售的桑蚕丝作为研究对象,分别用15 W和45 W的紫外灯进行人工加速老化,研究光处理对其结构和性能的影响。研究表明:光处理后,桑蚕丝纤维的结晶度随着光照时间的增加下降,光照处理均使其表面出现裂隙,甚至断裂,且45 W的紫外光老化程度更严重。     

Effects of Arginine Deimination and Citrulline Side-Chain Length on Peptide Secondary Structure Formation

Post-translational modifications expand the chemical functionality of peptides and proteins beyond that originating from the encoded amino acids, but studies on the structural effects of these modifications have been limited. Arginine undergoes deimination to give citrulline (Cit), converting the positively charged guanidinium moiety into a neutral urea group. Herein, we report the effect of Arg deimination on secondary structure formation. To understand the reason for the number of methylene units in Cit, the effect of Cit side-chain length on secondary structure formation was also studied. Ala-based peptides and β-hairpin peptides were used to study α-helix and β-sheet formation, respectively. Peptides containing Cit analogues were prepared by an orthogonal protecting group strategy coupled with solid-phase carbamylation. The CD data for the Ala-based peptides were analyzed by using modified Lifson–Roig theory, showing that the helix propensity of Arg decreased upon deimination and that either shortening or lengthening Cit also decreased the helix propensity. The β-hairpin peptides were analyzed by NMR methods, showing minimal change in strand formation energetics upon Arg deimination. Altering the Cit side-chain length did not affect strand formation energetics either. These results should be useful for the preparation of urea-bearing systems and the design of peptides incorporating urea-bearing residues with varying side-chain length.     

抗菌肽的结构功能关系及分子设计的研究现状

抗菌肽是一种广泛存于自然界中具有广谱抗菌作用的小分子多肽。文章首先对抗菌肽的结构进行分类、从肽链长度、α-螺旋结构、疏水性、两亲性和净电荷5个方面分析了分子结构对活性的影响、并对抗菌肽的分子设计进行了综述,对其发展前景进行了展望。     

含受阻胺结构的苯并三唑光稳定性和热稳定性研究

测试了4种2-[2’-羟基-3’-叔丁基-5’-（3-丙酸-4-哌啶酯基）苯基]-2H-苯并三唑结构类光稳定剂紫外吸收光谱、光稳定性及热稳定性。4种化合物在270～400nm均有较强的吸收峰,与引入受阻胺结构前苯并三唑光稳定剂相比．紫外吸收波长和摩尔消光系数不变。4种化合物的光稳定性和热稳定性明显高于引入受阻胺结构前的苯并三唑光稳定剂。光老化12h后,4种化合物仅分解3．2％～6．9％,引入受阻胺结构前苯并三唑则分解35．0％～36．3％。热稳定性测试表明。4种化合物失重1％时的温度为264～272℃．而引入受阻胺结构前苯并三唑失重1％时的温度分别为227℃和235℃。该类化合物分子内同时具有紫外吸收剂和自由基捕获剂两种功能,为高效光稳定剂。     

轻钢结构住宅体系及其发展

论述了轻钢结构住宅的优点及其常用结构体系,对设计和施工中存在的问题进行了探讨,阐述了轻钢结构住宅的推广思路和发展方向。     

The Protective Function and Modification of Secondary Metabolite Accumulation in Response to Light Stress in Dracocephalum forrestii Shoots

The aim of this work was to determine the effect of stress conditions caused by different light sources, i.e., blue LED (λ = 430 nm), red LED (λ = 670 nm), blue and red LED (70%:30%) and white LED (430–670 nm) on the growth and morphology of cultivated in vitro Dracocephalum forrestii shoot culture. It also examines the effects on bioactive phenolic compound production and photosynthetic pigment content, as well as on antioxidant enzyme activity (CAT, SOD, POD) and antioxidant properties. The most beneficial proliferation effect was observed under white LEDs (7.1 ± 2.1 shoots per explant). The white and blue lights stimulated the highest fresh weight gain, while red light induced the highest dry weight gain. The total phenolic acid content ranged from 13.824 ± 1.181 to 20.018 ± 801 mg g DW⁻¹ depending on light conditions. The highest content of rosmarinic acid was found in the control shoots (cultivated under fluorescent lamps), followed by culture grown under red light. All LED treatments, especially red and blue, increased salvianolic acid B content, and blue increased apigenin p-coumarylrhamnoside biosynthesis. The greatest ferric reduction activity was observed in shoots cultivated under red light, followed by blue; this is associated with the presence of the highest total phenol content, especially phenolic acids. Similarly, the highest DPPH radical scavenging potential was observed under red light followed by blue. This study proves that LEDs have emerged as significant support for directed in vitro propagation, taking advantage of specific stress responses on various light spectra. This study also showed how stress induced by different LED light spectra increases in Dracocephalum forrestii the synthesis of pharmacologically-active compounds. Hence, light stress may turn out to be a simpler alternative to metabolic engineering for improving the production of secondary metabolites of therapeutic value.