400-998-5282
專注多肽 服務(wù)科研
作為 CCK 的拮抗劑,PP 抑制胰腺的沉淀。PP 還作用于胃腸蠕動(dòng)、食物攝入和新陳代謝。
編號(hào):158392
CAS號(hào):1816258-36-2
單字母:Biotinyl-APLEPVYPGDNATPEQMAQYAADLRRYINMLTRPRY-CONH2
編號(hào): | 158392 |
中文名稱: | 胰腺多肽Biotin-PancreaticPoly Peptide , human |
英文名: | Biotin-PancreaticPoly Peptide , human |
CAS號(hào): | 1816258-36-2 |
單字母: | Biotinyl-APLEPVYPGDNATPEQMAQYAADLRRYINMLTRPRY-CONH2 |
三字母: | Biotinyl N端生物素標(biāo)記 -Ala丙氨酸 -Pro脯氨酸 -Leu亮氨酸 -Glu谷氨酸 -Pro脯氨酸 -Val纈氨酸 -Tyr酪氨酸 -Pro脯氨酸 -Gly甘氨酸 -Asp天冬氨酸 -Asn天冬酰胺 -Ala丙氨酸 -Thr蘇氨酸 -Pro脯氨酸 -Glu谷氨酸 -Gln谷氨酰胺 -Met甲硫氨酸 -Ala丙氨酸 -Gln谷氨酰胺 -Tyr酪氨酸 -Ala丙氨酸 -Ala丙氨酸 -Asp天冬氨酸 -Leu亮氨酸 -Arg精氨酸 -Arg精氨酸 -Tyr酪氨酸 -Ile異亮氨酸 -Asn天冬酰胺 -Met甲硫氨酸 -Leu亮氨酸 -Thr蘇氨酸 -Arg精氨酸 -Pro脯氨酸 -Arg精氨酸 -Tyr酪氨酸 -CONH2C端酰胺化 |
氨基酸個(gè)數(shù): | 36 |
分子式: | C195H301N55O56S3 |
平均分子量: | 4408.01 |
精確分子量: | 4405.16 |
等電點(diǎn)(PI): | 11.31 |
pH=7.0時(shí)的凈電荷數(shù): | 4 |
平均親水性: | -0.11333333333333 |
疏水性值: | -0.74 |
外觀與性狀: | 白色粉末狀固體 |
消光系數(shù): | 5960 |
來(lái)源: | 人工化學(xué)合成,僅限科學(xué)研究使用,不得用于人體。 |
純度: | 95%、98% |
鹽體系: | 可選TFA、HAc、HCl或其它 |
生成周期: | 2-3周 |
儲(chǔ)存條件: | 負(fù)80℃至負(fù)20℃ |
標(biāo)簽: | 生物素標(biāo)記肽(Biotinyl) 胰腺多肽 |
As antagonist of CCK, PP inhibits pancreatic sedretion. PP also acts on the gastrointestinal motility, food intake, and metabolism.
專肽生物合成用于蛋白質(zhì)-蛋白質(zhì)相互作用研究的生物素化肽。盡管生物素可以在 N 端或 C 端引入(通過(guò)賴氨酸殘基),但我們建議使用 N 端修飾,因?yàn)樗杀镜?、成功率高、周轉(zhuǎn)時(shí)間短且易于操作。因?yàn)槎嚯暮铣墒菑?C 端到 N 端合成的,因此,N 端修飾是 SPPS步驟的最后一步,不需要額外的特定縮合步驟。相比之下,C 端修飾需要額外的步驟,并且通常更復(fù)雜。當(dāng)然,原則上生物素可以定位在任何地方。
生物素可以通過(guò)多種不同的接頭或間隔物與肽分離。盡管如此,還是建議包含一個(gè)靈活的間隔物,例如 Ahx(一個(gè) 6 碳接頭),以使生物素標(biāo)簽更加穩(wěn)定或靈活。
專肽生物在 N 端或 C 端提供生物素化:生物素-N 端、賴氨酸-生物素-肽中間和賴氨酸-生物素-C 端。
專肽生物還可以使用 Ahx 接頭或長(zhǎng)碳 (LC) 接頭提供生物素化:生物素-Ahx-N 末端、Lys-Ahx-生物素-肽中間、Lys-Ahx-生物素-C-末端。
(生物素結(jié)構(gòu))
示例:
GRGDS在N端和C端標(biāo)記生物素的結(jié)構(gòu)展示。
1、GRGDS在N端標(biāo)記生物素,不增加Ahx 接頭
2、GRGDS在N端標(biāo)記生物素,增加一個(gè)Ahx 接頭
3、GRGDS在C端標(biāo)記生物素,不增加Ahx 接頭
4、GRGDS在C端標(biāo)記生物素,增加一個(gè)Ahx 接頭。
DOI | 名稱 | |
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10.1038/sj.bjp.0704604 | Neuropeptide Y, Y1, Y2 and Y4 receptors mediate Y agonist responses in isolated human colon mucosa | 下載 |
10.1210/jc.2003-030630 | Pancreatic polypeptide reduces appetite and food intake in humans | 下載 |
10.1002/ijc.20948 | Neuropeptide Y receptors in renal cell carcinomas and nephroblastomas | 下載 |
10.1038/oby.2006.180 | Physiological evidence for the involvement of peptide YY in the regulation of energy homeostasis in humans | 下載 |
10.1016/j.peptides.2012.06.008 | Surface masking shapes the traffic of the neuropeptide Y Y2 receptor | 下載 |
10.1002/oby.20534 | PYY3-36 and pancreatic polypeptide reduce food intake in an additive manner via distinct hypothalamic dependent pathways in mice | 下載 |
10.3390/ijms15034856 | Dimers of G-protein coupled receptors as versatile storage and response units | 下載 |
10.1021/acs.jmedchem.6b00310 | Optically Pure, Structural, and Fluorescent Analogues of a Dimeric Y4 Receptor Agonist Derived by an Olefin Metathesis Approach | 下載 |
10.1021/acs.bioconjchem.6b00376 | Identification of a Cyanine-Dye Labeled Peptidic Ligand for Y1R and Y4R, Based upon the Neuropeptide Y C-Terminal Analogue, BVD-15 | 下載 |
10.1016/0167-0115(93)90435-b | Proteolytic processing of neuropeptide Y and peptide YY by dipeptidyl peptidase IV | 下載 |
10.1371/journal.pone.0120346 | Cytotoxic helix-rich oligomer formation by melittin and pancreatic polypeptide | 下載 |
多肽Biotin-Ala-Pro-Leu-Glu-Pro-Val-Tyr-Pro-Gly-Asp-Asn-Ala-Thr-Pro-Glu-Gln-Met-Ala-Gln-Tyr-Ala-Ala-Asp-Leu-Arg-Arg-Tyr-Ile-Asn-Met-Leu-Thr-Arg-Pro-Arg-Tyr-NH2的合成步驟:
1、合成MBHA樹(shù)脂:取若干克MBHA樹(shù)脂(如初始取代度為0.5mmol/g)和1倍樹(shù)脂摩爾量的Fmoc-Linker-OH加入到反應(yīng)器中,加入DMF,攪拌使氨基酸完全溶解。再加入樹(shù)脂2倍量的DIEPA,攪拌混合均勻。再加入樹(shù)脂0.95倍量的HBTU,攪拌混合均勻。反應(yīng)3-4小時(shí)后,用DMF洗滌3次。用2倍樹(shù)脂體積的10%乙酸酐/DMF 進(jìn)行封端30分鐘。然后再用DMF洗滌3次,甲醇洗滌2次,DCM洗滌2次,再用甲醇洗滌2次。真空干燥12小時(shí)以上,得到干燥的樹(shù)脂{Fmoc-Linker-MHBA Resin},測(cè)定取代度。這里測(cè)得取代度為 0.3mmol/g。結(jié)構(gòu)如下圖:
2、脫Fmoc:取2.1g的上述樹(shù)脂,用DCM或DMF溶脹20分鐘。用DMF洗滌2遍。加3倍樹(shù)脂體積的20%Pip/DMF溶液,鼓氮?dú)?0分鐘,然后2倍樹(shù)脂體積的DMF 洗滌5次。得到 H2N-Linker-MBHA Resin 。(此步驟脫除Fmoc基團(tuán),茚三酮檢測(cè)為藍(lán)色,Pip為哌啶)。結(jié)構(gòu)圖如下:
3、縮合:取1.89mmol Fmoc-Tyr(tBu)-OH 氨基酸,加入到上述樹(shù)脂里,加適當(dāng)DMF溶解氨基酸,再依次加入3.78mmol DIPEA,1.8mmol HBTU。反應(yīng)30分鐘后,取小樣洗滌,茚三酮檢測(cè)為無(wú)色。用2倍樹(shù)脂體積的DMF 洗滌3次樹(shù)脂。(洗滌樹(shù)脂,去掉殘留溶劑,為下一步反應(yīng)做準(zhǔn)備)。得到Fmoc-Tyr(tBu)-Linker-MBHA Resin。氨基酸:DIPEA:HBTU:樹(shù)脂=3:6:2.85:1(摩爾比)。結(jié)構(gòu)圖如下:
4、依次循環(huán)步驟二、步驟三,依次得到
H2N-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Pro-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
以上中間結(jié)構(gòu),均可在專肽生物多肽計(jì)算器-多肽結(jié)構(gòu)計(jì)算器中,一鍵畫出。
最后再經(jīng)過(guò)步驟二得到 H2N-Ala-Pro-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin,結(jié)構(gòu)如下:
5、生物素反應(yīng)連接:在上述樹(shù)脂中,加入適當(dāng)DMF后,再加入1.89mmol 生物素到樹(shù)脂中,再加入3.78mmol DIPEA、1.8mmol HBTU,鼓氮?dú)夥磻?yīng)30分鐘。用2倍樹(shù)脂體積的DMF 洗滌3次樹(shù)脂(洗滌樹(shù)脂,去掉殘留溶劑,為下一步反應(yīng)做準(zhǔn)備)。 得到Biotin-Ala-Pro-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHAResin。 結(jié)構(gòu)如下:
5、切割:6倍樹(shù)脂體積的切割液(或每1g樹(shù)脂加8ml左右的切割液),搖床搖晃 2小時(shí),過(guò)濾掉樹(shù)脂,用冰無(wú)水乙醚沉淀濾液,并用冰無(wú)水乙醚洗滌沉淀物3次,最后將沉淀物放真空干燥釜中,常溫干燥24小試,得到粗品Biotin-Ala-Pro-Leu-Glu-Pro-Val-Tyr-Pro-Gly-Asp-Asn-Ala-Thr-Pro-Glu-Gln-Met-Ala-Gln-Tyr-Ala-Ala-Asp-Leu-Arg-Arg-Tyr-Ile-Asn-Met-Leu-Thr-Arg-Pro-Arg-Tyr-NH2。結(jié)構(gòu)圖見(jiàn)產(chǎn)品結(jié)構(gòu)圖。
切割液選擇: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%
(前兩種適合沒(méi)有容易氧化的氨基酸,例如Trp、Cys、Met。第三種適合幾乎所有的序列。)
6、純化凍干:使用液相色譜純化,收集目標(biāo)峰液體,進(jìn)行凍干,獲得蓬松的粉末狀固體多肽。不過(guò)這時(shí)要取小樣復(fù)測(cè)下純度 是否目標(biāo)純度。
7、最后總結(jié):
杭州專肽生物技術(shù)有限公司(ALLPEPTIDE http://amynixphotography.com)主營(yíng)定制多肽合成業(yè)務(wù),提供各類長(zhǎng)肽,短肽,環(huán)肽,提供各類修飾肽,如:熒光標(biāo)記修飾(CY3、CY5、CY5.5、CY7、FAM、FITC、Rhodamine B、TAMRA等),功能基團(tuán)修飾肽(疊氮、炔基、DBCO、DOTA、NOTA等),同位素標(biāo)記肽(N15、C13),訂書肽(Stapled Peptide),脂肪酸修飾肽(Pal、Myr、Ste),磷酸化修飾肽(P-Ser、P-Thr、P-Tyr),環(huán)肽(酰胺鍵環(huán)肽、一對(duì)或者多對(duì)二硫鍵環(huán)),生物素標(biāo)記肽,PEG修飾肽,甲基化修飾肽等。
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