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Fluorogenic substrate for dipeptidyl peptidase IV (DPP IV) and Xaa-Pro dipeptidase. Substrate for fibroblast activation protein alpha (FAP, also called antiplasmin-cleaving enzyme, APCE).
編號:181847
CAS號:125791-92-6
單字母:H2N-AP-AFC
編號: | 181847 |
中文名稱: | 二肽Fluorogenic DPP IV substrate; APCE substrate |
中文同義詞: | 二肽基肽酶IV(DPP IV)底物 |
CAS號: | 125791-92-6 |
單字母: | H2N-AP-AFC |
三字母: | H2N N端氨基 -Ala丙氨酸 -Pro脯氨酸 -AFC7-氨基-4-三氟甲基香豆素 |
氨基酸個(gè)數(shù): | 2 |
分子式: | C18H18N2O4F3 |
平均分子量: | 383.34 |
精確分子量: | 383.12 |
等電點(diǎn)(PI): | - |
pH=7.0時(shí)的凈電荷數(shù): | 0.97 |
平均親水性: | -0.5 |
疏水性值: | 0.1 |
外觀與性狀: | 白色粉末狀固體 |
消光系數(shù): | - |
來源: | 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 |
儲(chǔ)存條件: | 負(fù)80℃至負(fù)20℃ |
標(biāo)簽: | AFC修飾肽 二肽(Dipeptide) |
Fluorogenic substrate for dipeptidyl peptidase IV (DPP IV) and Xaa-Pro dipeptidase. Substrate for fibroblast activation protein alpha (FAP, also called antiplasmin-cleaving enzyme, APCE).
多肽熒光標(biāo)記由于沒有放射性,實(shí)驗(yàn)操作簡單。因此,目前在生物學(xué)研究中多肽熒光標(biāo)記應(yīng)用非常廣泛,多肽熒光標(biāo)記方法與熒光試劑的結(jié)構(gòu)有關(guān)系,對于有游離羧基的采用的方法與接多肽反應(yīng)相同,也采用HBTU/HOBt/DIEA方法連接。 在N端標(biāo)記FITC的多肽需經(jīng)歷環(huán)化作用來形成熒光素,通常會(huì)伴有最后一個(gè)氨基酸的去除,但當(dāng)有一個(gè)間隔器如氨基己酸,或者是通過非酸性環(huán)境將目的多肽從樹脂上切下來時(shí),這種情況可避免在切割的過程中被TFA切割掉。
人們利用利用熒光標(biāo)記的多肽來檢測目標(biāo)蛋白的活性,并將 其發(fā)展的高通量活性篩選方法應(yīng)用于疾病治療靶點(diǎn)蛋白的藥物篩選和藥物開發(fā)(例如,各種激 酶、磷酸酶、肽酶等)。
專肽生物能夠提供技術(shù)成熟的各種熒光標(biāo)記多肽。
下面是一些常見的多肽修飾熒光物質(zhì)結(jié)構(gòu):
FITC標(biāo)記
FITC(異硫氰酸熒光素)具有比較高的活性,我們公司可以通過兩種方式將FITC標(biāo)記于多肽 上:(1) 將FITC標(biāo)記于賴氨酸(Lys)或被選擇性地脫保護(hù)的鳥氨酸(ornithine)側(cè)鏈氨基 上;(2) 將FITC標(biāo)記于多肽N端氨基。
當(dāng)在N端標(biāo)記時(shí),建議在最后一個(gè)氨基和由異硫氰酸酯與氨基反應(yīng)產(chǎn)生的硫脲鍵之間引入 烷基間隔器(alkyl spacer),如氨基己酸(Ahx)。鏈接切割需要酸性環(huán)境,在N端標(biāo)記FITC 的多肽需經(jīng)歷環(huán)化作用來形成熒光素,通常會(huì)伴有最后一個(gè)氨基酸的去除,但當(dāng)有一個(gè)間隔器 如氨基己酸,或者是通過非酸性環(huán)境將目的肽從樹脂上切下來時(shí),這種情況可避免。空間位阻 被認(rèn)為是在熒光染料前使用Ahx的主要原因,而不是為什么FITC不能直接偶聯(lián)在多肽上的原因。
Ahx或b-Ala均可作為間隔器用于FITC標(biāo)記的多肽上。
普通熒光修飾
熒光修飾中文名稱 | N端 | N端帶有l(wèi)inker |
生物素標(biāo)記多肽 | Biotin- | Biotin-Ahx- |
異硫氰酸熒光素 | FITC- | FITC-Ahx- |
5-羧基熒光素 | 5-FAM- | 5-FAM-Ahx- |
丹磺酰熒光素 | Dansyl- | Dansyl-Ahx- |
5-羧基四甲基羅丹明 | TMR- (TAMRA-) | TMR-Ahx- (TAMRA-Ahx-) |
多肽N端 | 多肽序列中間 | N端帶有l(wèi)inker |
生物素標(biāo)記多肽 | Biotin- | 多肽C端 |
Lys(Biotin)- | -Lys(Biotin)-- | -Lys(Biotin) |
Lys(FITC)- | -Lys(FITC)- | -Lys(FITC) |
Lys(5-FAM)- | -Lys(5-FAM)- | -Lys(5-FAM) |
Lys(Dansyl)- | -Lys(Dansyl)- | -Lys(Dansyl) |
Lys(TMR)- | -Lys(TMR)- | -Lys(TMR) |
Lys(Dnp)- | -Lys(Dnp)- | -Lys(Dnp) |
專肽常做的熒光物質(zhì)的激發(fā)光波長和發(fā)射光波長??晒﹨⒖歼x擇:
熒光基團(tuán) | Ex(nm) | Em(nm) | 熒光基團(tuán) | Ex(nm) | Em(nm) |
羥基香豆素 | 325 | 386 | R-phycoerythrin (PE) (489) | 565 | 578 |
丹磺酰氯 | 340 | 578 | Rhodamine Red-X | 560 | 580 |
AMC | 345 | 445 | Tamara | 565 | 580 |
甲氧基香豆素 | 360 | 410 | Alexa fluor 555 | 556 | 573 |
Alexa fluor 系列 | 345 | 442 | Alexa fluor 546 | 556 | 573 |
氨基香豆素 | 350 | 445 | Rox | 575 | 602 |
Dabcyl | 453 | - | Alexa fluor 568 | 578 | 603 |
Cy2 | 490 | 510 | Texas Red | 589 | 615 |
FAM | 495 | 517 | Alexa fluor 594 | 590 | 617 |
Alexa fluor 488 | 494 | 517 | Alexa fluor | 621 | 639 |
FITC | 495 | 519 | Alexa fluor 633 | 650 | 668 |
Alexa fluor 430 | 430 | 545 | Cy5 (625) | 650 | 670 |
5-FAM | 492 | 518 | Alexa fluor 660 | 663 | 690 |
Alexa fluor 532 | 530 | 530 | Cy5.5 | 675 | 694 |
HEX | 535 | 556 | TruRed | 490; 675 | 695 |
5-TAMRA | 542 | 568 | Alexa fluor 680 | 679 | 702 |
Cy3 | 550 | 570 | Cy7 | 743 | 767 |
TRITC | 547 | 572 | Cy3.5 | 581 | 596 |
DOI | 名稱 | |
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10.1006/prep.2001.1572 | Expression, purification, and kinetic characterization of full-length human fibroblast activation protein | 下載 |
10.1042/BJ20021643 | Catalytic properties and inhibition of proline-specific dipeptidyl peptidases II, IV and VII | 下載 |
10.1021/bi026846s | Structural requirements for catalysis, expression, and dimerization in the CD26/DPIV gene family | 下載 |
10.1110/ps.03352504 | N-linked glycosylation of dipeptidyl peptidase IV (CD26): effects on enzyme activity, homodimer formation, and adenosine deaminase binding | 下載 |
10.1110/ps.03460604 | Crystal structure of human dipeptidyl peptidase IV in complex with a decapeptide reveals details on substrate specificity and tetrahedral intermediate formation | 下載 |
10.1016/j.bbaexp.2004.03.010 | Dipeptidyl peptidase 9 has two forms, a broad tissue distribution, cytoplasmic localization and DPIV-like peptidase activity | 下載 |
10.1111/j.1742-4658.2006.05253.x | Extraenzymatic functions of the dipeptidyl peptidase IV-related proteins DP8 and DP9 in cell adhesion, migration and apoptosis | 下載 |
10.1021/bi701921b | Fibroblast activation protein peptide substrates identified from human collagen I derived gelatin cleavage sites | 下載 |
10.1016/j.ejphar.2008.04.047 | Pharmacokinetic, pharmacodynamic, and efficacy profiles of alogliptin, a novel inhibitor of dipeptidyl peptidase-4, in rats, dogs, and monkeys | 下載 |
10.1002/art.27395 | Inhibition of fibroblast activation protein and dipeptidylpeptidase 4 increases cartilage invasion by rheumatoid arthritis synovial fibroblasts | 下載 |
10.1002/pro.443 | The dimeric transmembrane domain of prolyl dipeptidase DPP-IV contributes to its quaternary structure and enzymatic activities | 下載 |
10.1371/journal.pone.0038744 | Linagliptin improves insulin sensitivity and hepatic steatosis in diet-induced obesity | 下載 |
10.1210/jc.2013-1029 | The DPP-4 inhibitor linagliptin restores β-cell function and survival in human isolated islets through GLP-1 stabilization | 下載 |
10.1016/j.fob.2013.12.001 | Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs | 下載 |
10.1038/srep14421 | A whole-cell tumor vaccine modified to express fibroblast activation protein induces antitumor immunity against both tumor cells and cancer-associated fibroblasts | 下載 |
10.1111/bph.13549 | Glucagon-like peptide-1 receptor signalling reduces microvascular thrombosis, nitro-oxidative stress and platelet activation in endotoxaemic mice | 下載 |
10.1046/j.1432-1327.1998.2540650.x | Mouse fibroblast-activation protein--conserved Fap gene organization and biochemical function as a serine protease | 下載 |
10.1016/j.bbapap.2005.09.013 | Molecular characterization of a novel dipeptidyl peptidase like 2-short form (DPL2-s) that is highly expressed in the brain and lacks dipeptidyl peptidase activity | 下載 |
10.1021/acs.jmedchem.6b00475 | Comparative Analysis of Binding Kinetics and Thermodynamics of Dipeptidyl Peptidase-4 Inhibitors and Their Relationship to Structure | 下載 |