编号：200264
CAS号：1872441-22-9
单字母：H2N-YDEYLKQVIEVLETDPHFREKLQKADIEEI-OH
| 编号: | 200264 |
| 中文名称: | Nesfatin蛋白片段 Nesfatin-1 (30-59) (mouse, rat) |
| CAS号: | 1872441-22-9 |
| 单字母: | H2N-YDEYLKQVIEVLETDPHFREKLQKADIEEI-OH |
| 三字母: | H2N N端氨基 -Tyr酪氨酸 -Asp天冬氨酸 -Glu谷氨酸 -Tyr酪氨酸 -Leu亮氨酸 -Lys赖氨酸 -Gln谷氨酰胺 -Val缬氨酸 -Ile异亮氨酸 -Glu谷氨酸 -Val缬氨酸 -Leu亮氨酸 -Glu谷氨酸 -Thr苏氨酸 -Asp天冬氨酸 -Pro脯氨酸 -His组氨酸 -Phe苯丙氨酸 -Arg精氨酸 -Glu谷氨酸 -Lys赖氨酸 -Leu亮氨酸 -Gln谷氨酰胺 -Lys赖氨酸 -Ala丙氨酸 -Asp天冬氨酸 -Ile异亮氨酸 -Glu谷氨酸 -Glu谷氨酸 -Ile异亮氨酸 -OHC端羧基 |
| 氨基酸个数: | 30 |
| 分子式: | C167H260N40O54 |
| 平均分子量: | 3692.09 |
| 精确分子量: | 3689.88 |
| 等电点(PI): | 4.94 |
| pH=7.0时的净电荷数: | -1.78 |
| 平均亲水性: | 0.51379310344828 |
| 疏水性值: | -0.8 |
| 消光系数: | 2980 |
| 来源: | 人工化学合成，仅限科学研究使用，不得用于人体。 |
| 储存条件: | 负80℃至负20℃ |
| 标签: | 垂体和下丘脑激素 |
| DOI | 名称 | |
|---|---|---|
| 10.1038/nature05162 | Identification of nesfatin-1 as a satiety molecule in the hypothalamus | 下载 |
| 10.1038/nm1106-1248 | Keeping the fat off with nesfatin | 下载 |
| 10.1016/j.peptides.2010.09.027 | Central nesfatin-1 reduces the nocturnal food intake in mice by reducing meal size and increasing inter-meal intervals | 下载 |
| 10.1016/j.peptides.2012.03.015 | Nesfatin-1(30-59) but not the N- and C-terminal fragments, nesfatin-1(1-29) and nesfatin-1(60-82) injected intracerebroventricularly decreases dark phase food intake by increasing inter-meal intervals in mice | 下载 |
多肽H2N-Tyr-Asp-Glu-Tyr-Leu-Lys-Gln-Val-Ile-Glu-Val-Leu-Glu-Thr-Asp-Pro-His-Phe-Arg-Glu-Lys-Leu-Gln-Lys-Ala-Asp-Ile-Glu-Glu-Ile-COOH的合成步骤：
1、合成CTC树脂：称取2.18g CTC Resin（如初始取代度约为0.75mmol/g）和1.96mmol Fmoc-Ile-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4.91mmol 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-Ile-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-Ile-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4.91mmol Fmoc-Glu(OtBu)-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9.81mmol DIPEA，4.66mmol 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-Glu(OtBu)-Ile-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-Glu(OtBu)-Ile-CTC Resin
Fmoc-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Lys(Boc)-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Lys(Boc)-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Leu-Lys(Boc)-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Leu-Lys(Boc)-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Tyr(tBu)-Leu-Lys(Boc)-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Tyr(tBu)-Leu-Lys(Boc)-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Glu(OtBu)-Tyr(tBu)-Leu-Lys(Boc)-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Glu(OtBu)-Tyr(tBu)-Leu-Lys(Boc)-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Asp(OtBu)-Glu(OtBu)-Tyr(tBu)-Leu-Lys(Boc)-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
H2N-Asp(OtBu)-Glu(OtBu)-Tyr(tBu)-Leu-Lys(Boc)-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC Resin
Fmoc-Tyr(tBu)-Asp(OtBu)-Glu(OtBu)-Tyr(tBu)-Leu-Lys(Boc)-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-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-Tyr(tBu)-Asp(OtBu)-Glu(OtBu)-Tyr(tBu)-Leu-Lys(Boc)-Gln(Trt)-Val-Ile-Glu(OtBu)-Val-Leu-Glu(OtBu)-Thr(tBu)-Asp(OtBu)-Pro-His(Trt)-Phe-Arg(Pbf)-Glu(OtBu)-Lys(Boc)-Leu-Gln(Trt)-Lys(Boc)-Ala-Asp(OtBu)-Ile-Glu(OtBu)-Glu(OtBu)-Ile-CTC 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-Tyr-Asp-Glu-Tyr-Leu-Lys-Gln-Val-Ile-Glu-Val-Leu-Glu-Thr-Asp-Pro-His-Phe-Arg-Glu-Lys-Leu-Gln-Lys-Ala-Asp-Ile-Glu-Glu-Ile-COOH。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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