专肽生物_定制多肽合成服务_多肽合成定制服务_多肽合成厂家_多肽合成公司

一种软体动物心脏兴奋性神经肽，有可能用于细胞生物学、药理学和蛋白质相互作用研究。本产品为FMRF酰胺。

A molluscan cardioexcitatory neuropeptide which has the potential to be used in cell biology, pharmacological, and protein-protein interaction studies. This product is available as an FMRF-Amide.

软体动物神经肽心脏兴奋性和冻干固体，与神经肽Y一起定位在大脑的某些区域。该肽还抑制心脏肌膜囊泡中的Na+-Ca2+交换。

Molluscan neuropeptide cardioexcitatory and lyophilized solid that localizes with neuropeptide Y in some regions of the brain. This peptide also inhibits Na+-Ca2+ exchange in cardiac sarcolemmal vesicles.

FMRF 酰胺是经典的神经肽基序，广泛用于无脊椎动物神经肌肉与调节信号研究。其短序列支持 β 转角形成的高分辨分析。研究者通过分析芳香族残基与碱性残基的相互作用来确定结合决定因素。该分子有助于 RFamide 家族结构生物学研究。

FMRF-amide is a classical neuropeptide motif widely used to study invertebrate neuromuscular and modulatory signaling. Its short sequence supports high-resolution analyses of β-turn formation. Researchers examine aromatic and basic residue interplay to define binding determinants. The molecule contributes to RFamide-family structural biology.

FMRF amide的定义

FMRFamide是一种软体动物神经活性肽，由于直接激活阿米洛利敏感的钠通道1，可诱导快速兴奋性去极化反应【1】。

FMRFamide is a molluscan neuroactive peptide which induces a fast excitatory depolarizing response due to direct activation of amiloride-sensitive sodium channels【1】.

FMRF amide的相关肽

FLRFamide，LFRFamide，FFRFamide，LLRFamide，D-FMRFamide是FMRFamide类似物，已被发现与FMRFamide表现出交叉相互作用。这些肽也可能与激动剂作用于同一类RFamide受体，引起交叉脱敏【2】。两种无脊椎动物神经肽类似物IPPQFMRF酰胺（IF-8酰胺）和EGDEDEFLRF酰胺（EF-10酰胺），分别来自两种不同物种的非洲金丝桃蛙，Kassina maculata和Phylitimantis verrucosus的防御皮肤分泌物，代表了来自脊椎动物来源的第一个经典FMRF酰胺相关肽（FARP）【3】。

FLRFamide, LFRFamide, FFRFamide, LLRFamide, D-FMRFamide, are FMRFamide analogs that have been found to exhibit a cross-interaction with FMRFamide.  It is possible that these peptides also act on the same class of RFamide receptors as agonists to cause cross desensitization【2】. Two invertebrate neuropeptide analogues, IPPQFMRF amide (IF-8 amide) and EGDEDEFLRF amide (EF-10 amide), from the defensive skin secretions of two different species of African hyperoliid frogs, Kassina maculata and Phylictimantis verrucosus, respectively, represent the first canonical FMRF amide-related peptides (FaRPs) from a vertebrate source【3】.

 FMRF amide的发现

DA Price和MJ Greenberg于1977年从蛤蜊Macrocallista nimbosa中分离出第一种FMRF酰胺作为心脏兴奋性分子【4】。

The first FMRF-amide was isolated in 1977 as a cardioexcitatory molecule from the clam Macrocallista nimbosa, by DA Price and MJ Greenberg【4】.

 FMRF amide的结构特征

它是一种四肽神经递质，是与FLRFamide相同的RFamide肽家族的成员，具有相同的C端RFamide序列。FMRFamide的结构首先通过Edman dansyl降解和胰蛋白酶消化的组合确定，并通过合成证实【5】。

It is a tetrapeptide neurotransmitter, a member of the same family of RFamide peptides as FLRFamide, sharing the same C terminal RFamide sequence. The structure of FMRFamide was first determined by the combined use of Edman dansyl degradation and tryptic digestion and confirmed by synthesis【5】.

 Structure-activity relations (SAR) of FMRFamide on the isolated Rapana heart5 have shown that:

(1) The C-terminal RFamide is critical for activity; potency is markedly diminished by substitution with D amino acids and is abolished upon removal of the amide.

(2) The N-terminal phenylalanine and the methionine could be replaced by other residues, but a total length of at least four residues is important for activity.

(3) N-terminal elongation may have little effect.

(4) FMRFamide was the most potent of 14 peptides tested【6】.

 FMRF amide的作用方式

据报道，FMRFamides通过直接激活FMRFamide门控钠通道而不涉及G蛋白来发挥其作用。然而，早期对软体动物的电生理研究表明，FMRFamide也可以激活GPCR【6】。

It has been reported that FMRFamides exert their effect by directly activating FMRFamide-gated sodium channels without involvement of a G protein. However, earlier electrophysiological studies in molluscs suggested that FMRFamide could also activate a GPCR【6】.

 FMRF amide的功能

FMRFamides在幼虫和成年中枢神经系统以及选定的外周靶标中充当神经递质/神经调节剂。后者包括与进食（肠道，唾液腺），生殖（辅助腺，精囊和输卵管），运动（骨骼肌），循环（主动脉）和蜕皮（内脏和骨骼肌的协调调节）相关的组织【6】。

FMRFamides act as neurotransmitters/neuromodulators within the larval and adult CNS, as well as at selected peripheral targets. The latter include, for example, tissues associated with feeding (gut, salivary glands), reproduction (accessory glands, spermatheca, and oviducts), movement (skeletal muscle), circulation (aorta), and ecdysis (coordinated modulation of visceral and skeletal muscles) 【6】.

 FMRF amide的相关文献 

1. Lingueglia E, Champigny G, Lazdunski M, Barbry P (1995). Cloning of the amiloride-sensitive FMRFamide peptide-gated sodium channel. Nature, 378(6558):730-733.

2. Chen ML, Sharma R, Walker RJ (1995). Structure-activity studies of RFamide analogues on central neurones of Helix aspersa. Regulatory peptides,58:99-105.

3. Wang L, Smyth A, Johnsen AH, Zhou M, Chen T, Walker B, Shaw C (2009). FMRFamide-related peptides (FaRPs): A new family of peptides from amphibian defensive skin secretions. BBRC,  383(3):314-319

4. Price DA, Greenberg MJ (1977). Structure of a molluscan cardioexcitatory neuropeptide. Science, 197(4304):670-671.

5.  Kobayashi M , Muneoka Y (1989). Functions, Receptors, and Mechanisms of the FMRFamide-Related Peptides. Biol. Bull, 177: 206-209.

6. Meeusen T, Mertens I, Clynen E, Baggerman G, Nichols R, Nachman RJ, Huybrechts R, De Loof A, Schoofs L (2002). Identification in Drosophila melanogaster of the invertebrate G protein-coupled FMRFamide receptor. PNAS, 99(24):15363-15368.

多肽H2N-Phe-Met-Arg-Phe-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1.05g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0.95mmol Fmoc-Phe-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1.89mmol DIPEA，0.9mmol 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-Phe-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-Phe-Linker-MBHA Resin

Fmoc-Arg(Pbf)-Phe-Linker-MBHA Resin

H2N-Arg(Pbf)-Phe-Linker-MBHA Resin

Fmoc-Met-Arg(Pbf)-Phe-Linker-MBHA Resin

H2N-Met-Arg(Pbf)-Phe-Linker-MBHA Resin

Fmoc-Phe-Met-Arg(Pbf)-Phe-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-Phe-Met-Arg(Pbf)-Phe-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-Phe-Met-Arg-Phe-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。

以上所有内容，为专肽生物原创内容，请勿发布到其他网站上。