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Stresscopin 相关肽是促肾上腺皮质激素释放激素受体 2 型 (CRHR2) 的特异性配体。转录物在大脑和多种外周组织中表达。腹膜内注射应激蛋白相关肽(人)可抑制麻醉大鼠的热诱导水肿形成。它还减少食物摄入量并对胃排空活动表现出抑制作用。这种 CRHR2 激动剂可能代表一种在压力后维持体内平衡的内源性配体。
编号:138419
CAS号:348626-74-4
单字母:H2N-HPGSRIVLSLDVPIGLLQILLEQARARAAREQATTNARILARV-CONH2
应激蛋白相关肽是促肾上腺皮质激素释放激素受体2型(CRHR2)的特异性配体。转录本在脑和多个外周组织中表达。腹腔注射应激肽相关肽(人)抑制麻醉大鼠热诱导水肿的形成。它还可以减少食物摄入量,并对胃排空活动有抑制作用。这种CRHR2激动剂可能是一种内源性配体,在应激后维持体内平衡。
Stresscopin-related peptide is a specific ligand for the corticotropin-releasing hormone receptor type 2 (CRHR2). Transcripts are expressed in brain and multiple peripheral tissues. Intraperitoneal injections of stresscopin-related peptide (human) suppresses heat-induced edema formation in anesthetized rats. It also decreases food intake and exhibits an inhibitory effect on gastric emptying activity. This CRHR2 agonist might represent an endogenous ligand for maintaining homeostasis after stress.
应激蛋白相关肽(Human)是一类多肽类蛋白质。该肽是糖皮质激素受体的激活剂,也可与其它受体结合。应激蛋白相关肽(人)已被证明与人体内的离子通道和配体(如抗体)相互作用。
Stresscopin-Related Peptide (Human) is a protein that belongs to the class of peptides. The peptide is an activator of receptor for glucocorticoids, and it also can bind with other receptors. Stresscopin-Related Peptide (Human) has been shown to interact with ion channels and ligands, such as antibodies, in the human body.
人应激相关肽含有促肾上腺皮质激素释放因子家族基序,支持螺旋构象与受体结合。序列结构揭示应激轴信号多样性。折叠动力学可与天然应激肽比较。研究人员利用其解析促肾上腺皮质激素释放因子受体调控的结构决定因素。
Stresscopin-Related Peptide, human, contains CRF-family motifs that support helical conformations and receptor binding. Sequence architecture provides insight into stress-axis signaling diversity. Folding dynamics allow comparison with native stresscopin peptides. Researchers use it to dissect structural determinants of CRF receptor modulation.
人应激相关肽(游离酸)来源于促肾上腺皮质激素释放因子家族,具有两亲性螺旋结构。游离酸末端保留 C 端天然电荷。其序列基序适用于受体结合与信号分析。研究人员将其用于应激轴多肽定位与结构‑功能研究。
Stresscopin-Related Peptide (free acid) (human) derives from the corticotropin-releasing factor family, featuring amphipathic helices. Free acid termination preserves native charge at the C-terminus. Sequence motifs support receptor-binding and signaling analyses. Researchers use it in stress-axis peptide mapping and structure–function studies.
| DOI | 名称 | |
|---|---|---|
| 10.1111/j.1471-4159.2007.05052.x | Expression, binding, and signaling properties of CRF2(a) receptors endogenously expressed in human retinoblastoma Y79 cells: passage-dependent regulation of functional receptors | 下载 |
多肽H2N-His-Pro-Gly-Ser-Arg-Ile-Val-Leu-Ser-Leu-Asp-Val-Pro-Ile-Gly-Leu-Leu-Gln-Ile-Leu-Leu-Glu-Gln-Ala-Arg-Ala-Arg-Ala-Ala-Arg-Glu-Gln-Ala-Thr-Thr-Asn-Ala-Arg-Ile-Leu-Ala-Arg-Val-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2.19g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1.97mmol Fmoc-Val-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3.94mmol DIPEA,1.87mmol 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-Val-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-Val-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Ile-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Ile-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Ile-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Arg(Pbf)-Ile-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Ser(tBu)-Arg(Pbf)-Ile-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Ser(tBu)-Arg(Pbf)-Ile-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Gly-Ser(tBu)-Arg(Pbf)-Ile-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Gly-Ser(tBu)-Arg(Pbf)-Ile-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-Pro-Gly-Ser(tBu)-Arg(Pbf)-Ile-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
H2N-Pro-Gly-Ser(tBu)-Arg(Pbf)-Ile-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-Linker-MBHA Resin
Fmoc-His(Trt)-Pro-Gly-Ser(tBu)-Arg(Pbf)-Ile-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-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-His(Trt)-Pro-Gly-Ser(tBu)-Arg(Pbf)-Ile-Val-Leu-Ser(tBu)-Leu-Asp(OtBu)-Val-Pro-Ile-Gly-Leu-Leu-Gln(Trt)-Ile-Leu-Leu-Glu(OtBu)-Gln(Trt)-Ala-Arg(Pbf)-Ala-Arg(Pbf)-Ala-Ala-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Thr(tBu)-Thr(tBu)-Asn(Trt)-Ala-Arg(Pbf)-Ile-Leu-Ala-Arg(Pbf)-Val-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-His-Pro-Gly-Ser-Arg-Ile-Val-Leu-Ser-Leu-Asp-Val-Pro-Ile-Gly-Leu-Leu-Gln-Ile-Leu-Leu-Glu-Gln-Ala-Arg-Ala-Arg-Ala-Ala-Arg-Glu-Gln-Ala-Thr-Thr-Asn-Ala-Arg-Ile-Leu-Ala-Arg-Val-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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