编号: | 199889 |
中文名称: | CART(1-39), Human, Rat |
英文名: | CART(1-39), Human, Rat |
单字母: | Pyr-EDAELQPRALDIYSAVDDASHEKELPRRQLRAPGAVLQ-OH |
三字母: | Pyr 焦谷氨酸 -Glu谷氨酸 -Asp天冬氨酸 -Ala丙氨酸 -Glu谷氨酸 -Leu亮氨酸 -Gln谷氨酰胺 -Pro脯氨酸 -Arg精氨酸 -Ala丙氨酸 -Leu亮氨酸 -Asp天冬氨酸 -Ile异亮氨酸 -Tyr酪氨酸 -Ser丝氨酸 -Ala丙氨酸 -Val缬氨酸 -Asp天冬氨酸 -Asp天冬氨酸 -Ala丙氨酸 -Ser丝氨酸 -His组氨酸 -Glu谷氨酸 -Lys赖氨酸 -Glu谷氨酸 -Leu亮氨酸 -Pro脯氨酸 -Arg精氨酸 -Arg精氨酸 -Gln谷氨酰胺 -Leu亮氨酸 -Arg精氨酸 -Ala丙氨酸 -Pro脯氨酸 -Gly甘氨酸 -Ala丙氨酸 -Val缬氨酸 -Leu亮氨酸 -Gln谷氨酰胺 -OHC端羧基 |
氨基酸个数: | 38 |
分子式: | C188H303N57O63 |
平均分子量: | 4369.76 |
精确分子量: | 4367.23 |
等电点(PI): | 4.85 |
pH=7.0时的净电荷数: | -1.76 |
平均亲水性: | 0.57352941176471 |
疏水性值: | -0.75 |
外观与性状: | 白色粉末状固体 |
消光系数: | 1490 |
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或其它 |
生成周期: | 2-3周 |
储存条件: | 负80℃至负20℃ |
标签: | CARTS |
Definition
Cocaine- and amphetamine- regulated transcript (CART) peptides, derived from proCART polypeptide in humans are neuropeptides expressed in the brain1. They mainly serve as inhibitors of food intake1.
Discovery
CART was first purified by gel filtration and sequenced in 1980 from rat hypothalamus2.
Classification
CART is a neuropeptide family member2.
Structural Characteristics
CART gene encodes a peptide of either 129 or 116 amino acid residues in rat whereas only the short form exists in humans. The predicted signal sequence is 27 amino acid residues resulting in a precursor of 102 or 89 residues3. The C-terminal end of CART, consisting of 48 amino acid residues and 3 disulphide bonds, is thought to constitute a biologically active part of the molecule. Various CART peptides can be generated from the precursor: CARTs (55-102), (85-102), (55-76) and (62-76)3. CART (55-102) contains three disulphide bridges that are required for its function4.
Mode of action
The mode of action of CART peptides has not been fully established. Several studies have observed that they may function through CART receptors in turn activating down stream signaling pathways5.
Functions
CART is distributed in the central nervous system and periphery and has many physiological roles. It elicits similar behavior as cocaine. CART has been shown to have variety of effects on dopamine6. It increases blood pressure, has variety of behavioral effects and influences nociception6. In the hypothalamus CART mainly regulates energy homeostasis6. CART is also an endogenous inhibitor of food intake6. CART peptides are also important in anxiety, pain, arousal, startle response, regulation of calcium channels, and neuroendocrine hormone secretion5.
References
1. Thim L, Kristensen P, Larsen PJ, Wulff BS (1998). CART, a new anorectic peptide. Int J Biochem Cell Biol, 30(12), 1281-4.
2. Spiess J and Vale W (1980). Multiple forms of somatostatin-like activity in rat hypothalamus. Biochemistry 19, 2861–66.
3. Dylag T, Kotlinska J, Rafalski P, Pachuta A, Silberring J (2006). The activity of CART peptide fragments. Peptides, 27(8), 1926-33.
4. Murphy KG (2005). Dissecting the role of cocaine and amphetamine-regulated transcript (CART) in the control of appetite. Briefing in Functional Genomics and proteomic, 4 (2), 95–111.
5. Rogge G, Jones D, Hubert GW, Lin Y & Kuhar MJ (2008). CART peptides: regulators of body weight, reward and other functions. Nature Reviews Neuroscience, 9, 747-758.
6. Vicentic A, Jones DC (2007). The CART (cocaine- and amphetamine-regulated transcript) system in appetite and drug addiction. J Pharmacol Exp Ther., 320(2), 499-506.
多肽Pyr-Glu-Asp-Ala-Glu-Leu-Gln-Pro-Arg-Ala-Leu-Asp-Ile-Tyr-Ser-Ala-Val-Asp-Asp-Ala-Ser-His-Glu-Lys-Glu-Leu-Pro-Arg-Arg-Gln-Leu-Arg-Ala-Pro-Gly-Ala-Val-Leu-Gln-COOH的合成步骤:
1、合成CTC树脂:称取1.26g CTC Resin(如初始取代度约为1.17mmol/g)和1.77mmol Fmoc-Gln(Trt)-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4.42mmol 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-Gln(Trt)-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-Gln(Trt)-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4.42mmol Fmoc-Leu-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8.85mmol DIPEA,4.2mmol 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-Leu-Gln(Trt)-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-Leu-Gln(Trt)-CTC Resin
Fmoc-Val-Leu-Gln(Trt)-CTC Resin
H2N-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Gln(Trt)-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Gln(Trt)-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Leu-Gln(Trt)-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Leu-Gln(Trt)-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Glu(OtBu)-Leu-Gln(Trt)-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Glu(OtBu)-Leu-Gln(Trt)-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Ala-Glu(OtBu)-Leu-Gln(Trt)-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Ala-Glu(OtBu)-Leu-Gln(Trt)-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Asp(OtBu)-Ala-Glu(OtBu)-Leu-Gln(Trt)-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
H2N-Asp(OtBu)-Ala-Glu(OtBu)-Leu-Gln(Trt)-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin
Fmoc-Glu(OtBu)-Asp(OtBu)-Ala-Glu(OtBu)-Leu-Gln(Trt)-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-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-Glu(OtBu)-Asp(OtBu)-Ala-Glu(OtBu)-Leu-Gln(Trt)-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTC Resin,结构如下:
5、焦谷氨酸反应连接:在上述树脂中,加入适当DMF后,再加入4.42mmol 焦谷氨酸到树脂中,再加入8.85mmol DIPEA、4.2mmol 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Pyr-Glu(OtBu)-Asp(OtBu)-Ala-Glu(OtBu)-Leu-Gln(Trt)-Pro-Arg(Pbf)-Ala-Leu-Asp(OtBu)-Ile-Tyr(tBu)-Ser(tBu)-Ala-Val-Asp(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-His(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Leu-Pro-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Leu-Arg(Pbf)-Ala-Pro-Gly-Ala-Val-Leu-Gln(Trt)-CTCResin。 结构如下:
6、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Pyr-Glu-Asp-Ala-Glu-Leu-Gln-Pro-Arg-Ala-Leu-Asp-Ile-Tyr-Ser-Ala-Val-Asp-Asp-Ala-Ser-His-Glu-Lys-Glu-Leu-Pro-Arg-Arg-Gln-Leu-Arg-Ala-Pro-Gly-Ala-Val-Leu-Gln-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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