Neuropeptide originally isolated from porcine upper small intestine.
编号：195446
CAS号：88813-36-9
单字母：H2N-GWTLNSAGYLLGPHAIDNHRSFHDKYGLA-CONH2
Neuropeptide originally isolated from porcine upper small intestine.
| DOI | 名称 | |
|---|---|---|
| 10.1016/j.autneu.2004.04.002 | Coexistence of non-adrenergic non-cholinergic inhibitory and excitatory neurotransmitters in a large neuronal subpopulation in the vaginal segment of the chicken oviduct | 下载 |
| 10.14670/HH-20.509 | Response of the gut neuroendocrine system of Leuciscus cephalus (L.) to the presence of Pomphorhynchus laevis Müller, 1776 (Acanthocephala) | 下载 |
| 10.3354/dao066245 | Histopathology, ultrastructure and immunohistochemistry of Coregonus lavaretus hearts naturally infected with Ichthyocotylurus erraticus (Trematoda) | 下载 |
| 10.1073/pnas.87.18.7105 | PD134308, a selective antagonist of cholecystokinin type B receptor, enhances the analgesic effect of morphine and synergistically interacts with intrathecal galanin to depress spinal nociceptive reflexes | 下载 |
| 10.1111/j.1365-2761.2007.00833.x | Selected pathological, immunohistochemical and ultrastructural changes associated with an infection by Diphyllobothrium dendriticum (Nitzsch, 1824) (Cestoda) plerocercoids in Coregonus lavaretus (L.) (Coregonidae) | 下载 |
| 10.1111/j.1365-2982.2010.01529.x | The chemical code of porcine enteric neurons and the number of enteric glial cells are altered by dietary probiotics | 下载 |
| 10.1002/cne.23131 | Galanin gene expression and effects of its knock-down on the development of the nervous system in larval zebrafish | 下载 |
| 10.1111/ahe.12179 | Galanin is Co-Expressed with Substance P, Calbindin and Corticotropin-Releasing Factor (CRF) in The Enteric Nervous System of the Wild Boar (Sus scrofa) Small Intestine | 下载 |
| 10.1016/0014-5793(83)80033-7 | Galanin - a novel biologically active peptide from porcine intestine | 下载 |
多肽H2N-Gly-Trp-Thr-Leu-Asn-Ser-Ala-Gly-Tyr-Leu-Leu-Gly-Pro-His-Ala-Ile-Asp-Asn-His-Arg-Ser-Phe-His-Asp-Lys-Tyr-Gly-Leu-Ala-NH2的合成步骤：
1、合成MBHA树脂：取若干克的MBHA树脂（如初始取代度为0.5mmol/g）和1倍树脂摩尔量的Fmoc-Linker-OH加入到反应器中，加入DMF，搅拌使氨基酸完全溶解。再加入树脂2倍量的DIEPA，搅拌混合均匀。再加入树脂0.95倍量的HBTU，搅拌混合均匀。反应3-4小时后，用DMF洗涤3次。用2倍树脂体积的10%乙酸酐/DMF 进行封端30分钟。然后再用DMF洗涤3次，甲醇洗涤2次，DCM洗涤2次，再用甲醇洗涤2次。真空干燥12小时以上，得到干燥的树脂{Fmoc-Linker-MHBA Resin}，测定取代度。这里测得取代度为 0.3mmol/g。结构如下图：
2、脱Fmoc：取2.45g的上述树脂，用DCM或DMF溶胀20分钟。用DMF洗涤2遍。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-Linker-MBHA 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2.21mmol Fmoc-Ala-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4.41mmol DIPEA，2.09mmol 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-Ala-Linker-MBHA 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-Ala-Linker-MBHA Resin
Fmoc-Leu-Ala-Linker-MBHA Resin
H2N-Leu-Ala-Linker-MBHA Resin
Fmoc-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Ser(tBu)-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Ser(tBu)-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Asn(Trt)-Ser(tBu)-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Asn(Trt)-Ser(tBu)-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Leu-Asn(Trt)-Ser(tBu)-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Leu-Asn(Trt)-Ser(tBu)-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Thr(tBu)-Leu-Asn(Trt)-Ser(tBu)-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Thr(tBu)-Leu-Asn(Trt)-Ser(tBu)-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Trp(Boc)-Thr(tBu)-Leu-Asn(Trt)-Ser(tBu)-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
H2N-Trp(Boc)-Thr(tBu)-Leu-Asn(Trt)-Ser(tBu)-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin
Fmoc-Gly-Trp(Boc)-Thr(tBu)-Leu-Asn(Trt)-Ser(tBu)-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA 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-Gly-Trp(Boc)-Thr(tBu)-Leu-Asn(Trt)-Ser(tBu)-Ala-Gly-Tyr(tBu)-Leu-Leu-Gly-Pro-His(Trt)-Ala-Ile-Asp(OtBu)-Asn(Trt)-His(Trt)-Arg(Pbf)-Ser(tBu)-Phe-His(Trt)-Asp(OtBu)-Lys(Boc)-Tyr(tBu)-Gly-Leu-Ala-Linker-MBHA Resin，结构如下：
5、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H2N-Gly-Trp-Thr-Leu-Asn-Ser-Ala-Gly-Tyr-Leu-Leu-Gly-Pro-His-Ala-Ile-Asp-Asn-His-Arg-Ser-Phe-His-Asp-Lys-Tyr-Gly-Leu-Ala-NH2。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。）
6、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。
7、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，甲基化修饰肽等。
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