An excellent, highly soluble and specific substrate for a routine assay of human and rat leukocyte and porcine pancreatic elastases. MeOSuc-AAPV-pNA is also cleaved by myeloblastin.
编号：181969
CAS号：70967-90-7
单字母：MeOSuc-AAPV-pNA
| 编号: | 181969 |
| 中文名称: | 四肽MeOSuc-Ala-Ala-Pro-Val-对硝基苯胺 |
| 中文同义词: | N-甲氧基琥珀酰-丙酰氨-丙酰氨-脯酰氨-缬氨酸对硝基酰苯胺 |
| CAS号: | 70967-90-7 |
| 单字母: | MeOSuc-AAPV-pNA |
| 三字母: | MeOSuc 暂无说明 -Ala丙氨酸 -Ala丙氨酸 -Pro脯氨酸 -Val缬氨酸 -pNA对硝基苯胺 |
| 氨基酸个数: | 4 |
| 分子式: | C27H38O9N6 |
| 平均分子量: | 590.63 |
| 精确分子量: | 590.27 |
| 等电点(PI): | - |
| pH=7.0时的净电荷数: | - |
| 平均亲水性: | -0.83333333333333 |
| 疏水性值: | 1.55 |
| 外观与性状: | 白色粉末状固体 |
| 消光系数: | - |
| 来源: | 人工化学合成，仅限科学研究使用，不得用于人体。 |
| 纯度: | 95%、98% |
| 盐体系: | 可选TFA、HAc、HCl |
| 储存条件: | 负80℃至负20℃ |
| 标签: | pNA修饰肽 |
An excellent, highly soluble and specific substrate for a routine assay of human and rat leukocyte and porcine pancreatic elastases. MeOSuc-AAPV-pNA is also cleaved by myeloblastin.
| DOI | 名称 | |
|---|---|---|
| 10.1021/bi015650+ | Heterogeneity in serpin-protease complexes as demonstrated by differences in the mechanism of complex breakdown | 下载 |
| 10.1046/j.1432-1033.2002.03316.x | Rodent alpha-chymases are elastase-like proteases | 下载 |
| 10.1074/jbc.M301512200 | Extended substrate specificity of rat mast cell protease 5, a rodent alpha-chymase with elastase-like primary specificity | 下载 |
| 10.1021/bi00152a011 | Changing the inhibitory specificity and function of the proteinase inhibitor eglin c by site-directed mutagenesis: functional and structural investigation | 下载 |
| 10.1164/rccm.200307-959OC | Effect of treatment with dornase alpha on airway inflammation in patients with cystic fibrosis | 下载 |
| 10.1016/S0014-5793(04)00187-5 | Killing of target cells by redirected granzyme B in the absence of perforin | 下载 |
| 10.1016/j.febslet.2005.08.056 | Differences between human proteinase 3 and neutrophil elastase and their murine homologues are relevant for murine model experiments | 下载 |
| 10.1007/s00253-006-0800-0 | Extremely high alkaline protease from a deep-subsurface bacterium, Alkaliphilus transvaalensis | 下载 |
| 10.1016/j.abb.2007.06.028 | Reversible inactivation of serpins at acidic pH | 下载 |
| 10.1007/s00253-007-1164-9 | Nucleotide and deduced amino acid sequences of a subtilisin-like serine protease from a deep-sea bacterium, Alkalimonas collagenimarina AC40(T) | 下载 |
| 10.1002/cbic.200800415 | New peptolides from the cyanobacterium Nostoc insulare as selective and potent inhibitors of human leukocyte elastase | 下载 |
| 10.3390/molecules14010378 | 2-Amino- and 2-alkylthio-4H-3,1-benzothiazin-4-ones: synthesis, interconversion and enzyme inhibitory activities | 下载 |
| 10.1111/j.1742-4658.2010.07663.x | Paradoxical interactions between modifiers and elastase-2 | 下载 |
| 10.1159/000327009 | Shear stress induces the release of an endothelial elastase: role in integrin α(v)β(3)-mediated FGF-2 release | 下载 |
| 10.1107/S1600536812038378 | Methyl 3-[(1,1-dioxo-1λ(6),2-benzothiazol-3-yl)amino]-5-nitrothiophene-2-carboxyl-ate | 下载 |
| 10.1016/j.str.2013.04.021 | A noncanonical mechanism of carboxypeptidase inhibition revealed by the crystal structure of the Tri-Kunitz SmCI in complex with human CPA4 | 下载 |
| 10.1016/s0006-291x(86)80262-5 | Comparison of peptide aldehydes with alpha 1-antitrypsin as elastase inhibitors for use in emphysema | 下载 |
| 10.1021/bi00379a016 | Catalysis by human leukocyte elastase: proton inventory as a mechanistic probe | 下载 |
| 10.1016/0006-291x(87)91350-7 | Reversible, slow, tight-binding inhibition of human leukocyte elastase | 下载 |
| 10.1016/0003-2697(85)90436-1 | Solid-phase radioimmunoassay for human neutrophil elastase: a sensitive method for determining secreted and cell-associated enzyme | 下载 |
| 10.1016/0003-2697(79)90043-5 | Sensitive substrates for human leukocyte and porcine pancreatic elastase: a study of the merits of various chromophoric and fluorogenic leaving groups in assays for serine proteases | 下载 |
| 10.1111/j.1432-1033.1984.tb08423.x | Similarities between human and rat leukocyte elastase and cathepsin G | 下载 |
| 10.1164/ajrccm.152.1.7599816 | Elastin and collagen degradation products in urine of patients with cystic fibrosis | 下载 |
| 10.1002/eji.1830241233 | Involvement of a serine protease in the synthesis of platelet-activating factor by endothelial cells stimulated by tumor necrosis factor-alpha or interleukin-1 alpha | 下载 |
| 10.1164/ajrccm.154.3.8810611 | Elastin-derived peptides and neutrophil elastase in bronchoalveolar lavage fluid | 下载 |
| 10.1183/09031936.98.11020377 | Catalase, myeloperoxidase and hydrogen peroxide in cystic fibrosis | 下载 |
| 10.1016/s0968-0896(00)00281-9 | 6-Acylamino-2-1[(ethylsulfonyl)oxy]-1H-isoindole-1,3-diones mechanism-based inhibitors of human leukocyte elastase and cathepsin G: effect of chirality in the 6-acylamino substituent on inhibitory potency and selectivity | 下载 |
| 10.1078/0944-7113-00031 | Anti-inflammatory and spasmolytic activity of extracts from Droserae herba | 下载 |
| 10.1016/j.molbiopara.2007.07.013 | Biochemical properties of an intracellular serpin from Echinococcus multilocularis | 下载 |
| 10.1021/ja807827k | Cleavage of four carbon-carbon bonds during biosynthesis of the griseorhodin a spiroketal pharmacophore | 下载 |
| 10.1021/je050303e | Systematic Measurements of Peptide Adsorption in Hydrophobic Chromatography | 下载 |
多肽MeOSuc-Ala-Ala-Pro-Val-pNA的合成步骤：
1、合成CTC树脂：称取1.42g CTC Resin（如初始取代度约为0.51mmol/g）和0.87mmol Fmoc-Val-OH于反应器中，加入适量DCM溶解氨基酸（需要注意，此时CTC树脂体积会增大好几倍，避免DCM溶液过少），再加入2.17mmol DIPEA（Mw：129.1,d：0.740g/ml），反应2-3小时后，可不抽滤溶液，直接加入1ml的HPLC级甲醇，封端半小时。依次用DMF洗涤2次，甲醇洗涤1次，DCM洗涤一次，甲醇洗涤一次，DCM洗涤一次，DMF洗涤2次（这里使用甲醇和DCM交替洗涤，是为了更好地去除其他溶质，有利于后续反应）。得到 Fmoc-Val-CTC Resin。结构图如下：
2、脱Fmoc：加3倍树脂体积的20%Pip/DMF溶液，鼓氮气30分钟，然后2倍树脂体积的DMF 洗涤5次。得到 H2N-Val-CTC Resin 。（此步骤脱除Fmoc基团，茚三酮检测为蓝色，Pip为哌啶）。结构图如下：
3、缩合：取2.17mmol Fmoc-Pro-OH 氨基酸，加入到上述树脂里，加适当DMF溶解氨基酸，再依次加入4.35mmol DIPEA，2.06mmol HBTU。反应30分钟后，取小样洗涤，茚三酮检测为无色。用2倍树脂体积的DMF 洗涤3次树脂。（洗涤树脂，去掉残留溶剂，为下一步反应做准备）。得到Fmoc-Pro-Val-CTC Resin。氨基酸：DIPEA：HBTU：树脂=3：6：2.85：1（摩尔比）。结构图如下：
4、依次循环步骤二、步骤三，依次得到
H2N-Pro-Val-CTC Resin
Fmoc-Ala-Pro-Val-CTC Resin
H2N-Ala-Pro-Val-CTC Resin
Fmoc-Ala-Ala-Pro-Val-CTC Resin
以上中间结构，均可在专肽生物多肽计算器-多肽结构计算器中，一键画出。
最后再经过步骤二得到 H2N-Ala-Ala-Pro-Val-CTC Resin，结构如下：
5、MeOSuc反应连接：在上述树脂中，加入适当DMF后，再加入2.17mmolMeOSuc到树脂中，再加入4.35mmol DIPEA，鼓氮气反应30分钟。用2倍树脂体积的DMF 洗涤3次树脂（洗涤树脂，去掉残留溶剂，为下一步反应做准备）。 得到MeOSuc-Ala-Ala-Pro-Val-CTC Resin。 结构如下：
6、全保护切割：配置0.5%TFA/DCM溶液，溶液体积约为树脂体积的3倍。再次用DCM洗涤树脂2遍（去除残留DMF），后将配置好的溶液倒入到反应器中，反应30分钟。抽滤树脂，收集滤液（此时多肽已经从树脂上分离，存在于滤液中）。多肽序列为 MeOSuc-Ala-Ala-Pro-Val-CTC Resin。 在滤液中添加DIEPA，调PH至7-8。用饱和NaHCO3洗涤滤液，分离出DCM层溶液。可适当旋蒸DCM层溶液，减少有机溶剂。再次加入1或2倍体积的乙酸乙酯，用稀HCl溶液调PH至微酸性，将多肽从DCM层萃取到乙酸乙酯层。用饱和NaCl洗涤2次乙酸乙酯层。用无水硫酸镁吸收乙酸乙酯层的水分。通过减压旋蒸，直接将乙酸乙酯完全旋蒸掉，得到晶体状固体多肽，用于下一步C端反应。或通过减压旋蒸保留适量乙酸乙酯的溶液体积，加入冰乙醚析出 多肽，然后对多肽进行烘干操作即可用于下一步C端反应。MeOSuc-Ala-Ala-Pro-Val-COOH的结构图如下。
7、4-硝基苯胺反应连接：在上述树脂中，加入适当DMF后，再加入2.17mmol 4-硝基苯胺到树脂中，再加入4.35mmol DIPEA、2.06mmol HBTU，鼓氮气反应30分钟。用2倍树脂体积的DMF 洗涤3次树脂（洗涤树脂，去掉残留溶剂，为下一步反应做准备）。 得到 MeOSuc-Ala-Ala-Pro-Val-pNA。 结构如下：
8、切割：6倍树脂体积的切割液（或每1g树脂加8ml左右的切割液），摇床摇晃 2小时，过滤掉树脂，用冰无水乙醚沉淀滤液，并用冰无水乙醚洗涤沉淀物3次，最后将沉淀物放真空干燥釜中，常温干燥24小试，得到粗品MeOSuc-Ala-Ala-Pro-Val-pNA。结构图见产品结构图。
切割液选择：1）TFA：H2O=95%：5%
2）TFA：H2O：TIS=95%：2.5%：2.5%
3）三氟乙酸：茴香硫醚：1，2-乙二硫醇：苯酚：水=87.5%：5%：2.5%：2.5%：2.5%
（前两种适合没有容易氧化的氨基酸，例如Trp、Cys、Met。第三种适合几乎所有的序列。）
9、纯化冻干：使用液相色谱纯化，收集目标峰液体，进行冻干，获得蓬松的粉末状固体多肽。不过这时要取小样复测下纯度 是否目标纯度。
10、最后总结：
杭州专肽生物技术有限公司（ALLPEPTIDE https://www.allpeptide.com）主营定制多肽合成业务，提供各类长肽，短肽，环肽，提供各类修饰肽，如：荧光标记修饰（CY3、CY5、CY5.5、CY7、FAM、FITC、Rhodamine B、TAMRA等），功能基团修饰肽（叠氮、炔基、DBCO、DOTA、NOTA等），同位素标记肽（N15、C13），订书肽（Stapled Peptide）,脂肪酸修饰肽（Pal、Myr、Ste），磷酸化修饰肽（P-Ser、P-Thr、P-Tyr），环肽（酰胺键环肽、一对或者多对二硫键环），生物素标记肽，PEG修饰肽，甲基化修饰肽
以上所有内容，为专肽生物原创内容，请勿发布到其他网站上。
