多肽RSMYEVQGHEMRLRV的參數(shù)和合成路線|三字母為Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val|專肽生物產(chǎn)品-定制多肽合成服務(wù)
文章目錄
一、多肽Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val|RSMYEVQGHEMRLRV的結(jié)構(gòu)圖
二、多肽Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val|RSMYEVQGHEMRLRV的多肽參數(shù)
三、多肽Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val|RSMYEVQGHEMRLRV的靜電荷圖
四、多肽Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val|RSMYEVQGHEMRLRV的親疏水比例統(tǒng)計(條形圖)
五、多肽Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val|RSMYEVQGHEMRLRV的親疏水性氨基酸、極性非極性氨基酸、酸堿性氨基酸的統(tǒng)計(扇形圖)
六、多肽Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val|RSMYEVQGHEMRLRV的合成步驟
專肽生物官網(wǎng):http://tsjxdd.com (以下圖片均由專肽生物多肽結(jié)構(gòu)計算器自動生成)
一、Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val的結(jié)構(gòu)圖如下
二、Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val的多肽參數(shù)如下
單字母:H2N-RSMYEVQGHEMRLRV-OH
三字母:H2N-Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val-COOH
中文序列:精氨酸-絲氨酸-甲硫氨酸-酪氨酸-谷氨酸-纈氨酸-谷氨酰胺-甘氨酸-組氨酸-谷氨酸-甲硫氨酸-精氨酸-亮氨酸-精氨酸-纈氨酸
氨基酸個數(shù):15
分子式:C79H131N27O23S2
平均分子量:1891.18
精確分子量(Exact Mass):1889.94
等電點(PI):11.06
pH=7.0時的凈電荷數(shù):2.21
GRAVY值:-0.89
親水殘基比例:-
消光系數(shù):1490
平均親水性及多肽溶解建議:親水(該多肽在純水中溶解性較好,大多數(shù)情況下無需助溶)
三、H2N-RSMYEVQGHEMRLRV-OH的靜電荷圖
四、多肽Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val的親疏水比例統(tǒng)計(條形圖)
五、多肽Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val的親疏水性氨基酸、極性非極性氨基酸、酸堿性氨基酸的統(tǒng)計(扇形圖)
六、多肽Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val的合成步驟:
1、合成CTC樹脂:稱取0.3g CTC Resin(如初始取代度約為1.0mmol/g)和0.4 mmol的Fmoc-Val-OH于反應(yīng)器中,加入適量DCM溶解氨基酸(需要注意,此時CTC樹脂體積會增大好幾倍,避免DCM溶液過少),再加入樹脂0.9 mmol的DIPEA((Mw:129.1,d:0.740g/ml)),反應(yīng)2-3小時后,可不抽濾溶液,直接加入1ml的HPLC級甲醇,封端半小時。依次用DMF洗滌2次,甲醇洗滌1次,DCM洗滌一次,甲醇洗滌一次,DCM洗滌一次,DMF洗滌2次(這里使用甲醇和DCM交替洗滌,是為了更好地去除其他溶質(zhì),有利于后續(xù)反應(yīng))。得到Fmoc-Val-CTC Resin備用。結(jié)構(gòu)圖如下:
2、脫Fmoc:加3倍樹脂體積的20%Pip/DMF溶液,鼓氮氣30分鐘,然后2倍樹脂體積的DMF 洗滌5次。得到H2N-Val-CTC Resin。(此步驟脫除Fmoc基團(tuán),茚三酮檢測為藍(lán)色)。結(jié)構(gòu)圖如下:
3、縮合:取2.7mmol Fmoc-Arg(Pbf)-OH 氨基酸,加入到上述樹脂里,加適當(dāng)DMF溶解氨基酸,再分別加入1.8mmol DIPEA,1.67mmol HBTU。反應(yīng)30分鐘后,用2倍樹脂體積的DMF 洗滌3次樹脂。(洗滌樹脂,去掉殘留溶劑,為下一步反應(yīng)做準(zhǔn)備)。得到Fmoc-Arg(Pbf)-Val-CTC Resin。氨基酸:DIPEA:HBTU:樹脂=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-Arg(Pbf)-Val-CTC Resin
Fmoc-Leu-Arg(Pbf)-Val-CTC Resin
H2N-Leu-Arg(Pbf)-Val-CTC Resin
Fmoc-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
H2N-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
Fmoc-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
H2N-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
Fmoc-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
H2N-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
Fmoc-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
H2N-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
Fmoc-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
H2N-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
Fmoc-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
H2N-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
Fmoc-Val-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
H2N-Val-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
Fmoc-Glu(OtBu)-Val-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
H2N-Glu(OtBu)-Val-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
Fmoc-Tyr(tBu)-Glu(OtBu)-Val-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
H2N-Tyr(tBu)-Glu(OtBu)-Val-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
Fmoc-Met-Tyr(tBu)-Glu(OtBu)-Val-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
H2N-Met-Tyr(tBu)-Glu(OtBu)-Val-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
Fmoc-Ser(tBu)-Met-Tyr(tBu)-Glu(OtBu)-Val-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
H2N-Ser(tBu)-Met-Tyr(tBu)-Glu(OtBu)-Val-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
Fmoc-Arg(Pbf)-Ser(tBu)-Met-Tyr(tBu)-Glu(OtBu)-Val-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin
最后再經(jīng)過步驟二得到 H2N-Arg(Pbf)-Ser(tBu)-Met-Tyr(tBu)-Glu(OtBu)-Val-Gln(Trt)-Gly-His(Trt)-Glu(OtBu)-Met-Arg(Pbf)-Leu-Arg(Pbf)-Val-CTC Resin,結(jié)構(gòu)圖如下:
5、切割:6倍樹脂體積的切割液(或每1g樹脂加8ml左右的切割液),搖床搖晃 2小時,過濾掉樹脂,用冰無水乙醚沉淀濾液,并用冰無水乙醚洗滌沉淀物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 Arg-Ser-Met-Tyr-Glu-Val-Gln-Gly-His-Glu-Met-Arg-Leu-Arg-Val。
切割液選擇:1)TFA:H2O=95%:5%、TFA:H2O=97.5%:2.5%
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。不過這時要取小樣復(fù)測下純度 是否目標(biāo)純度。
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