降钙素基因相关肽肽段的抗原性和免疫原性

用固相合成法合成了有关降钙素基因相关肽ＣＧＲＰ五个肽段。它们的氨基酸顺序分别为：ＣＧＲＰ－１（２４－３７）;ＣＧＲＰ－２（１４－３７）;ＣＧＲＰ－３（９－２３）;ＣＧＲＰ－４（１－２３）和ＣＧＲＰ－５（１－１３）。用酶标方法测定了它们的抗原性,发现ＣＧＲＰ－２和ＣＧＲＰ－１能够很好地与抗ＣＧＲＰ抗血清结合。将ＣＧＲＰ－１;ＣＧＲＰ－３和ＣＧＲＰ－５分别免疫兔子,获得的抗血清分别与ＣＧＲＰ进行反应,发现只有抗ＣＧＲＰ－１的抗血清能够与ＣＧＲＰ很好地结合。因此,可以推测ＣＧＲＰ的羧端部分是其免疫活性部位。     

降钙素基因相关肽肽段的抗原性和免疫原性

用固相合成法合成了有关降钙素基因相关肽ＣＧＲＰ五个肽段。它们的氨基酸顺序分别为：ＣＧＲＰ－１（２４－３７）;ＣＧＲＰ－２（１４－３７）;ＣＧＲＰ－３（９－２３）;ＣＧＲＰ－４（１－２３）和ＣＧＲＰ－５（１－１３）。用酶标方法测定了它们的抗原性,发现ＣＧＲＰ－２和ＣＧＲＰ－１能够很好地与抗ＣＧＲＰ抗血清结合。将ＣＧＲＰ－１;ＣＧＲＰ－３和ＣＧＲＰ－５分别免疫兔子,获得的抗血清分别与ＣＧＲＰ进行反应,     

降钙素基因相关肽的研究进展

降钙素基因相关肽（ＣＧＲＰ）是由３７个氨基酸残基构成的生物活性多肽,与降钙素（ＣＴ）源子一个共同的基因。ＣＧＲＰ分布广泛,具有很强的血管扩张、降低血压以及心肌正性肌力作用等,并参与心血管系统稳态的调节。目前,ＣＧＲＰ已能人工合成,将为某些心血管疾病如高血压、心肌缺血、痉挛性或闭塞性周围血管疾病等的治疗提供一条崭新的途径。     

降钙素基因相关肽的新近研究进展

降钙素基因相关肽（CGRP）是存在于感觉神经末梢的一种神经肽,是具有代表性的一种活性多肽。实验证明CGRP在身体许多组织器官起着重要的调节作用,特别是对脑具有很强的血管扩张、逆转血管痉挛、改善脑血液循环的作用。对肠道免疫功能也有调节作用,同时介导神经免疫系统的相互调节。     

化学合成的α—降钙素基因相关基因在大肠杆菌和哺乳…

通过化学法合成α－降钙素基因相关肽的基因,该基因５＇端有ＢａｍＨ Ｉ切点和启始密码,３＇端有ＤａｌＩ切点和终止密码。为了表达这一基因,组建表达载体ｐＸＺ６２．这一表达载体具有Ｔｒｅ启动子,含有Ａｐ＾ｒ和ＸｙＬＥ基因。α－降钙素基因相关肽基因与ｐＸＺ６２边接,构建重组质粒ｐＸＺ１０１;并将该基因与质粒ｐＨβＡＰｒ－３Ｐ－ｎｅｏ连接,构建重组质粒ｐＸＺ２０１．将重组质粒分别转化到大肠矸菌和ＨｅＬａ细胞     

降钙素和降钙素基因相关肽的选择性表达

降钙素和降钙素基因相关肽(CT/CGRP)由同一基因编码,该基因结构及其5＇端侧翼序列决定了它能够在甲状腺C细胞以及中枢和外周神经细胞生成不同的表达产物.这种选择表达调控决定多细胞生物的发育、性别分化和进化.如果表达失控将导致甲状腺髓样瘤(MTC)和骨质疏松症等疾病.文章对该基因的结构和选择性表达调控进行了综述.     

Simultaneous Post-cysteine(S-Acm) Group Removal Quenching of Iodine and Isolation of Peptide by One Step Ether Precipitation

The S-acetamidomethyl (Acm) protecting group is widely used in the chemical synthesis of peptides that contain one or more disulfide bonds. Treatment of peptides containing S-Acm protecting group with iodine results in simultaneous removal of the sulfhydryl protecting group and disulfide formation. However, the excess iodine needs to be quenched or adsorbed as quickly as possible after completion of the disulfide bond formation in order to minimize side reactions that are often associated with the iodination step. We report a simple method for simultaneous post-cysteine (Acm) group removal quenching of iodination and isolation. Use of large volumes of diethyl ether for direct precipitation action of the oxidized peptide from the 90 or 95% aqueous acetic acid solution affords nearly quantitative recovery of largely iodine-free peptide ready for direct purification. It was successfully applied to the synthesis of various peptides including human insulin-like peptide 3 analogues. Although recovery yields were comparable to the traditionally used ascorbic acid quenching method, this new approach offers significant advantages such as more simple utility, minimal side reactions, and greater cost effectiveness.     

降钙素基因相关肽（CGRP）受体的纯化和鉴定

降钙素基因相关肽（ＣＧＲＰ）为３７肽,它在体内主要分布于神经系统和心血管系统,是较强的扩张血管物质。人们已认识到脑部的ＣＧＲＰ最多,由于新鲜猪脑来源丰富,故我们选用猪脑为材料来纯化ＣＧＲＰ受体。以期为该受体的深入研究提供足够的材料和奠定基础,纯化的受体将用于制备抗ＣＧＲＰ受体的单克隆抗体,为了从猪脑中纯化ＣＲＲＰ受体,首先用含有胆盐的磷酸盐缓冲液将该受体从猪脑细胞膜上解离下来,解离下来的受体量较多,而杂蛋白较少,且受体活性可保持较长时间不降低。以离子交换柱层析（ＱＡＥ－Ｓｅｐｈａｄｅｘ－Ａ－２５）进行初步分离纯化,再用化学合成的ＣＧＲＰ与活化的琼脂糖（Ａｆｆｉ－Ｇｅｌ－１０,Ｂｉｏ－Ｒａｄ公司产品）制备的亲和层析柱进行亲和层析,取得的纯化受体保留了与１２５Ｉ－ＣａＧＲＰ结合的能力,而与降钙素（Ｃａｌｃｉｔｏｎｉｎ）神经肽Ｙ（ＮｅｕｒｏｐｅｐｔｉｄｅＹ）和Ｋａｔａｃａｌｃｉｎ等无交叉反应,但与有４６％氨基酸序列同源性的Ａｍｙｌｉｎ有一定交叉反应。纯化的受体经ＳＤＳ－聚丙烯酰胺凝胶电泳和高压液相色谱（ＨＰＬＣ）鉴定呈现单一蛋白条带或蛋白峰,其位置对应分子量为６６ｋＤ。     

The Synthesis of Phosphopeptides Using Microwave-assisted Solid Phase Peptide Synthesis

A series of phosphorylated peptides were synthesised using microwave mediated solid phase peptide synthesis. Acidic cleavage of peptides from the solid support using microwave irradiation often resulted in reattachment of the phosphate benzyl protecting group to the peptide chain. However for most phosphopeptide sequences performing the cleavage reaction at room temperature in order to minimize this undesired alkylation was successful. Notably for phosphopeptides containing a methionine residue flanking the phosphorylated residue (for serine and threonine) the trifluoroacetic acid mediated cleavage afforded the benzylated side product as a major component. This detrimental process was not observed for a corresponding tyrosine containing sequence.     

人降钙素基因相关肽转基因马铃薯的RT-PCR分析

报道经过农杆菌介导将人降钙素基因相关肽(calcitoningenerelatedpeptide,CGRP)基因由马铃薯块茎专一表达classIpatatin基因5′侧翼区和CaMV35S启动子驱动构建的马铃薯表达载体导入马铃薯,PCR鉴定获得了转基因植株。RTPCR分析证实classIpatatin基因5′侧翼区驱动的CGRPmRNA在转基因马铃薯中的表达。研究结果在开发转基因马铃薯生物反应器表达医用多肽中具有重要意义。     

致炎剂联合降钙素基因相关肽对大鼠硬脑膜神经源性炎症的影响

目的：探讨致炎剂联合降钙素基因相关肽对大鼠硬脑膜神经源性炎症的影响,寻求最佳慢性偏头痛的实验模型。方法：24只SD雄性大鼠。随机分为生理盐水组（NS）、降钙素基因相关肽组1（CGRe1,10-3M）、降钙素基因相关肽组2（CGRP2,10-4M）,以及IS（20μL）＋CGRP（10μL,10-4M）组,每组各6只,通过甲苯胺蓝染色法,观察肥大细胞脱颗粒,采用伊文氏蓝荧光显像法,观察大鼠硬脑膜血管渗出,采用多普勒激光血流仪检测法,观察各组硬脑膜动脉血流量变化。结果：与NS组相比,CGRPl、CGRP2、IS＋CGRP组肥大细胞脱颗粒数和比率、脑血流量均明显升高,P〈0．05,并且硬脑膜荧光红斑明显增多;与CGRPl、CGRP2组相比,IS＋CGRP组肥大细胞脱颗粒数和比率、脑血流量均明显升高,P〈0．05,并且硬脑膜荧光红斑明显增多。CGRP两组相比,上述指标比较,差异没有统计学意义,P〉0．05。结论：IS＋CGRP能够明显刺激大鼠硬脑膜发生神经源性炎症反应,可以作为慢性偏头痛动物实验模型。     

用混合氨基酸接肽的方法合成OX型肽亚库

肽库合成是药物研究组合化学策略的重要技术之一。建立合成OX型肽亚库的固相合成方法 ,接肽反应采用等摩尔的Fmoc氨基酸混合物和DIC HOBt缩合方法 ,以高浓度的缩合剂和不断缩减溶剂的策略促进偶联反应进行完全。产物的氨基酸组成分析结果显示 ,所使用的常见氨基酸都能以相近的摩尔比例接肽至X位置。推测经多次接肽后最终形成的肽亚库中 ,含低活力氨基酸较多的肽其浓度虽然会较低一些 ,但影响不会太大 ,且本合成方法成本相对较低 ,故可为抗原表位分析、多肽药物筛选及构效关系分析提供一种有用的工具。     

The engineering of membrane-permeable peptides

Reversible lipid attachment was investigated as a means to deliver small peptides into cells. Two labile straight chain alkyl motifs were developed: a cysteine dodecane disulfide (Cdd) building block and a tyrosine- or serine-myristate ester. Both moieties are cleaved on cell internalization and are compatible with Fmoc solid phase peptide synthesis. A series of fluorophore-labeled peptides that varied in lipophilic content, net charge, and charge distribution were synthesized. The peptides were screened for cellular uptake efficiency as monitored by fluorescence microscopy. Effective peptide transport is based on a distributed net positive charge introduced as lysine residues at the C and/or N terminus of the peptide and the presence of a hydrophobic domain exhibiting an estimated log P4.0. The incorporation of labile lipid motifs into peptides enhances lipophilic character of the peptides and contributes to cellular uptake with minimal alteration to the native sequence.     

肾血管性高血压大鼠模型中ET1和CGRP的变化

目的探讨降钙素基因相关肽（CGRP）及血浆内皮素（ET1）在高血压病中的作用。方法选用正常血压大鼠42只,随机分对照组、手术组、假手术组,分别观察血压值、CGRP及ET1值。结果手术组与正常组比较血压明显高于正常对照组（P〈0．01）,假手术组与正常对照组比较,血压无明显变化（P〉0．05）;手术组与对照组比较CGRP显著升高（P〈0．01）;假手术组与正常对照组比较,CGRP未见升高（P〉0．05）。手术组与对照组比较ET1无明显变化（P〉0．05）。结论CGRP在肾血管性高血压的发生发展中具有保护作用。ET1与CGRP是心血管系统中的一对拮抗因子,与CGRP的作用相反,ET1有较强的缩血管作用,从而导致血压升高。     

A NOVEL APPROACH TO SOLID PHASE CHEMICAL SYNTHESIS OF OLIGONUCLEOTIDE mRNA CAP ANALOGS

A novel approach for the synthesis of 5′-capped 2′-O-methyloligoribonucleotides on a disulfide-tethered solid support is described. The key step of the synthesis is ZnCl₂ promoted coupling of m⁷GDP imidazolide to a fully deprotected oligonucleotide 5′-phosphate on-support. By this methodology m⁷G⁵′pppm²′Apm²′Upm^2′Ap has been prepared.