400-998-5282
专注多肽 服务科研
400-998-5282
专注多肽 服务科研
PHI-27（猪源）是 27 残基猪肽组氨酸异亮氨酰胺，与 VIP 和相关肽结构相似。其两亲螺旋区域支持受体结合分析。研究人员将其用于胃肠道和神经内分泌信号传导研究。该肽有助于 PHI 家族结构的比较评估。
PHI-27 (porcine) is the 27-residue porcine peptide histidine isoleucinamide, structurally similar to VIP and related peptides. Its amphipathic helical region supports receptor-binding analyses. Researchers employ it in gastrointestinal and neuroendocrine signaling studies. The peptide aids comparative evaluation of PHI-family structures.
猪组氨酸异亮氨酸酰胺肽 (PHI)(1-27) 是一种胃肠神经肽，其特征性 N 端组氨酸对受体结合至关重要。其序列支持螺旋 - 转角结构及与血管活性肠肽 / 垂体腺苷酸环化酶激活肽系统交叉对话的分析。研究人员用其比较猪调控肽。该分子有助于定位肽介导的分泌通路。
Peptide Histidine Isoleucinamide (PHI), Porcine (1-27) is a gastrointestinal neuropeptide with a characteristic N-terminal histidine crucial for receptor binding. Its sequence supports analyses of helix–turn structure and cross-talk with VIP/PACAP systems. Researchers employ it to compare porcine regulatory peptides. The molecule aids mapping of peptide-mediated secretory pathways.
| DOI | 名称 | |
|---|---|---|
| 10.1016/0143-4179(88)90069-8 | Effects of peptide HI on basal and stimulated insulin and glucagon secretion in the mouse | 下载 |
| 10.1073/pnas.78.11.6603 | Isolation and characterization of the intestinal peptide porcine PHI (PHI-27), a new member of the glucagon--secretin family | 下载 |
多肽H2N-His-Ala-Asp-Gly-Val-Phe-Thr-Ser-Asp-Phe-Ser-Arg-Leu-Leu-Gly-Gln-Leu-Ser-Ala-Lys-Lys-Tyr-Leu-Glu-Ser-Leu-Ile-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2.73g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2.46mmol Fmoc-Ile-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4.91mmol DIPEA，2.33mmol 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-Ile-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-Ile-Linker-MBHA Resin
Fmoc-Leu-Ile-Linker-MBHA Resin
H2N-Leu-Ile-Linker-MBHA Resin
Fmoc-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Val-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Val-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Gly-Val-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Gly-Val-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Gly-Val-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Asp(OtBu)-Gly-Val-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-Ala-Asp(OtBu)-Gly-Val-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
H2N-Ala-Asp(OtBu)-Gly-Val-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-Linker-MBHA Resin
Fmoc-His(Trt)-Ala-Asp(OtBu)-Gly-Val-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-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)-Ala-Asp(OtBu)-Gly-Val-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Phe-Ser(tBu)-Arg(Pbf)-Leu-Leu-Gly-Gln(Trt)-Leu-Ser(tBu)-Ala-Lys(Boc)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Leu-Ile-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-Ala-Asp-Gly-Val-Phe-Thr-Ser-Asp-Phe-Ser-Arg-Leu-Leu-Gly-Gln-Leu-Ser-Ala-Lys-Lys-Tyr-Leu-Glu-Ser-Leu-Ile-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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