多肽修飾：N 端、內部和 C 端

N-端、內部和C-端多肽修飾對各種應用都很有用，例如Western印圖、蛋白質-蛋白質互作研究和基於螢光的檢測。請使用下表查看各種應用的清單，並跳轉到更多資訊，包括結構和相關參考文獻。

若要索取定制多肽的報價或尋求設計定制 PEPscreen® 多肽庫的幫助，請按以下連結。

          

多肽修飾	應用
乙醯化	藉由防止 N 端降解來增加多肽的穩定性
Dansyl and Fluorescein and 7-methoxycoumarin acetic acid	Protein-protein interaction and localization studies<
棕櫚酸	增加其細胞滲透性並幫助肽與細胞膜結合
環化 (二硫橋)	穩定多肽的構象，增加生物活性及酵素的穩定性
半胱氨酸氨甲基化 (CAM)	多肽質量指紋分析，用於識別和表徵多肽。在蛋白質測試中阻斷半胱氨酸殘基氧化
磷酸化	植物及動物的基因表達、蛋白質與蛋白質之間的相互作用及訊號傳導
同位素標示氨基酸同位素標示的氨基酸	研究蛋白質互動、蛋白質、轉譯後的修飾，如泛素化和磷酸化
減少肽與其負載結合部位的靜電阻礙

 

1.0 N-端修飾

1.1  乙醯化

這種修飾可移除肽 N 端上的正電荷，從而模仿天然蛋白質。在某些情況下，它可以防止 N 端降解，從而增加肽的穩定性 ^1-2

Molecular Weight	43 g/mol
Availability：	PEPscreen®

參考資料

1. Arnesen T. Towards a Functional Understanding of Protein N-Terminal Acetylation. PLoS Biol. 9(5):e1001074. https://doi.org/10.1371/journal.pbio.1001074

2. Wallace RJ. 1992. Acetylation of peptides inhibits their degradation by rumen micro-organisms. Br J Nutr. 68(2):365-372. https://doi.org/10.1079/bjn19920095

1.2  生物素

生物素對鏈霉親和素和avidin有很強的親和力。生物素標記的肽常用於免疫分析¹、組織細胞化學²和基於螢光的流式細胞計³

。

Molecular Weight	340 g/mol
Availability：	PEPscreen®

參考資料

1. Sélo I, Négroni L, Créminon C, Grassi J, Wal J. 1996. Preferential labeling of ?-amino N-terminal groups in peptides by biotin: application to the detection of specific anti-peptide antibodies by enzyme immunoassays. Journal of Immunological Methods. 199(2):127-138. https://doi.org/10.1016/s0022-1759(96)00173-1

2. HOWL J, WANG X, KIRK CJ, WHEATLEY M. 1993. Fluorescent and biotinylated linear peptides as selective bifunctional ligands for the V1a vasopressin receptor. Eur J Biochem. 213(2):711-719. https://doi.org/10.1111/j.1432-1033.1993.tb17811.x

3. Buranda, et. al. 1991. Peptide, antibodies and FRET on beads in flow cytometry: a model system using fluoresceinated and biotinylated β-endorphin. Cytometry . 3721-31.

1.3  Dansyl

Dansyl標記的縮氨酸用於基於螢光的檢測。

分子重量	249 g/mol
激發波長	372 nm
可用性：	PEPscreen®

參考資料

1. Glukhov E, Stark M, Burrows LL, Deber CM. 2005. Basis for Selectivity of Cationic Antimicrobial Peptides for BacterialVersusMammalian Membranes. J. Biol. Chem.. 280(40):33960-33967. https://doi.org/10.1074/jbc.m507042200

2. Matsuzaki K, Mitani Y, Akada K, Murase O, Yoneyama S, Zasloff M, Miyajima K. 1998. Mechanism of Synergism between Antimicrobial Peptides Magainin 2 and PGLa?. Biochemistry. 37(43):15144-15153. https://doi.org/10.1021/bi9811617

3. Pecht I, Maron E, Arnon R, Sela M. 1971. Specific Excitation Energy Transfer from Antibodies to Dansyl-Labeled Antigen. Studies with the "Loop" Peptide of Hen Egg-White Lysozyme. Eur J Biochem. 19(3):368-371. https://doi.org/10.1111/j.1432-1033.1971.tb01325.x

1.4 2，4-二硝基苯基

2，4-DNP 用作 (7-methoxy coumarin-4-yl) 乙酰 (MCA) 的淬火劑，有時也用作色氨酸 (tryptophan) 的淬火劑。這種修飾可以附著在肽的 N 端，或透過賴氨酸側鏈作為內部修飾。

分子重量	167 g/mol
激發波長	354-400 nm
可用性：	PEPscreen®

參考資料

1. Vickers C, Hales P, Kaushik V, Dick L, Gavin J, Tang J, Godbout K, Parsons T, Baronas E, Hsieh F, et al. 2002. Hydrolysis of Biological Peptides by Human Angiotensin-converting Enzyme-related Carboxypeptidase. J. Biol. Chem.. 277(17):14838-14843. https://doi.org/10.1074/jbc.m200581200

2. Knight C, Willenbrock F, Murphy G. 1992. A novel coumarin-labelled peptide for sensitive continuous assays of the matrix metalloproteinases. 296(3):263-266. https://doi.org/10.1016/0014-5793(92)80300-6

1.5  Fluorescein

螢光素標籤的肽有許多基於螢光素的生物分子應用，包括蛋白質-蛋白質互作、流式細胞計量和定位研究^1-3.

。

分子重量	359 g/mol
激發/發射波長	494/518 nm
可用性：	PEPscreen®

參考資料

1. Richard JP, Melikov K, Vives E, Ramos C, Verbeure B, Gait MJ, Chernomordik LV, Lebleu B. 2003. Cell-penetrating Peptides. J. Biol. Chem.. 278(1):585-590. https://doi.org/10.1074/jbc.m209548200

2. Farley RA, Tran CM, Carilli CT, Hawke D, Shively JE. 1984. The amino acid sequence of a fluorescein-labeled peptide from the active site of (Na,K)-ATPase. J Biol Chem. 259(15):9532-5.

3. Futaki S, Suzuki T, Ohashi W, Yagami T, Tanaka S, Ueda K, Sugiura Y. 2001. Arginine-rich Peptides. J. Biol. Chem.. 276(8):5836-5840. https://doi.org/10.1074/jbc.m007540200

4. Foerg C, Weller KM, Rechsteiner H, Nielsen HM, Fernández-Carneado J, Brunisholz R, Giralt E, Merkle HP. 2008. Metabolic Cleavage and Translocation Efficiency of Selected Cell Penetrating Peptides: A Comparative Study with Epithelial Cell Cultures. AAPS J. 10(2):349-359. https://doi.org/10.1208/s12248-008-9029-4

1.6  7-methoxycoumarin acetic acid (Mca)

7-methoxy coumarin-labeled peptides have applications in protein-protein interaction and localization studies^1-3．

分子重量	217 g/mol
激發/發射波長	323/382 nm
可用性：	PEPscreen®

參考資料

1. Vidal, et. al. 1996. Solid-Phase Synthesis and Cellular Localization of a C-and/or N-terminal Labeled Peptide. Journal of Peptide Science. 2125-133.

2. Yandek LE, Pokorny A, Florén A, Knoelke K, Langel Ü, Almeida PF. 2007. Mechanism of the Cell-Penetrating Peptide Transportan 10 Permeation of Lipid Bilayers. Biophysical Journal. 92(7):2434-2444. https://doi.org/10.1529/biophysj.106.100198

1.7  棕櫚酸

棕櫚酸是一種 16 碳脂肪酸，與多肽結合可增加其細胞滲透性，並有助於多肽與細胞膜的結合 ^1-2.

。

Molecular Weight	239 g/mol
Availability：	PEPscreen®

參考資料

1. Avrahami D, Shai Y. 2004. A New Group of Antifungal and Antibacterial Lipopeptides Derived from Non-membrane Active Peptides Conjugated to Palmitic Acid. J. Biol. Chem.. 279(13):12277-12285. https://doi.org/10.1074/jbc.m312260200

2. Buss JE, Sefton BM. 1986. Direct identification of palmitic acid as the lipid attached to p21ras.. Mol. Cell. Biol.. 6(1):116-122. https://doi.org/10.1128/mcb.6.1.116

分子量	N/A
Availability：	PEPscreen®、AQUA™多肽

參考資料

1. Góngora-Benítez M, Tulla-Puche J, Albericio F. 2014. Multifaceted Roles of Disulfide Bonds. Peptides as Therapeutics. Chem. Rev.. 114(2):901-926. https://doi.org/10.1021/cr400031z

2. Schmelz EA, Huffaker A, Carroll MJ, Alborn HT, Ali JG, Teal PE. 2012. An Amino Acid Substitution Inhibits Specialist Herbivore Production of an Antagonist Effector and Recovers Insect-Induced Plant Defenses. Plant Physiol.. 160(3):1468-1478. https://doi.org/10.1104/pp.112.201061

2.2  半胱氨酸氨甲酰化 (CAM)

氨甲酰化 (CAM) 是通過與碘乙酰胺反應引入半胱氨酸殘基的一種特意的轉譯後修飾。具有這種修飾的肽主要用於肽質量指紋分析（Peptide Mass Fingerprinting），以識別和表徵蛋白質¹。在其他檢測中，這個過程被用來阻止半胱氨酸被氧化²。

Molecular Weight	160 g/mol
Availability：	PEPscreen®、AQUA™多肽

參考資料

1. Wilkins MR, Appel RD, Williams KL, Hochstrasser DF. 2007. Proteome Research. https://doi.org/10.1007/978-3-540-72910-5

2. Rombouts I, Lagrain B, Brunnbauer M, Delcour JA, Koehler P. 2013. Improved identification of wheat gluten proteins through alkylation of cysteine residues and peptide-based mass spectrometry. Sci Rep. 3(1): https://doi.org/10.1038/srep02279

2.3  同位素標示的氨基酸

AQUA肽是具有富含 ¹⁸O、 ¹³C和/或 ¹⁴N的氨基酸的合成肽。它們在化學、物理特性和生物活性¹方面與原生肽類似。這些縮氨酸的主要應用是研究蛋白互作、蛋白質、轉譯後的修改，例如泛素化和磷酸化^2-5。

參考資料

1. Kettenbach AN, Rush J, Gerber SA. 2011. Absolute quantification of protein and post-translational modification abundance with stable isotope?labeled synthetic peptides. Nat Protoc. 6(2):175-186. https://doi.org/10.1038/nprot.2010.196

2. Li H, Xu C, Blais S, Wan Q, Zhang H, Landry SJ, Hioe CE. 2009. Proximal Glycans Outside of the Epitopes Regulate the Presentation of HIV-1 Envelope gp120 Helper Epitopes. J Immunol. 182(10):6369-6378. https://doi.org/10.4049/jimmunol.0804287

3. Le Bihan T, Grima R, Martin S, Forster T, Le Bihan Y. 2010. Quantitative analysis of low-abundance peptides in HeLa cell cytoplasm by targeted liquid chromatography/mass spectrometry and stable isotope dilution: emphasising the distinction between peptide detection and peptide identification. Rapid Commun. Mass Spectrom.. 24(7):1093-1104. https://doi.org/10.1002/rcm.4487

4. Santhoshkumar P, Raju M, Sharma KK. ?A-Crystallin Peptide 66SDRDKFVIFLDVKHF80 Accumulating in Aging Lens Impairs the Function of ?-Crystallin and Induces Lens Protein Aggregation. PLoS ONE. 6(4):e19291. https://doi.org/10.1371/journal.pone.0019291

5. Sato Y, Miyashita A, Iwatsubo T, Usui T. 2012. Simultaneous Absolute Protein Quantification of Carboxylesterases 1 and 2 in Human Liver Tissue Fractions using Liquid Chromatography-Tandem Mass Spectrometry. Drug Metab Dispos. 40(7):1389-1396. https://doi.org/10.1124/dmd.112.045054

6. Brun V, Masselon C, Garin J, Dupuis A. 2009. Isotope dilution strategies for absolute quantitative proteomics. Journal of Proteomics. 72(5):740-749. https://doi.org/10.1016/j.jprot.2009.03.007

2.4  磷酸化

磷酸化可在 Tyr、Ser 和 Thr 殘基上進行，作為多肽的一種翻譯後修飾 (PTM)。磷酸化的肽在許多細胞過程中都有應用，例如植物和動物的基因表達、蛋白質與蛋白質之間的互作和信號傳導^1,2。

Molecular Weight	82 g/mol
Availability：	PEPscreen®、AQUA™多肽

參考資料

1. Guenther JF, Chanmanivone N, Galetovic MP, Wallace IS, Cobb JA, Roberts DM. 2003. Phosphorylation of Soybean Nodulin 26 on Serine 262 Enhances Water Permeability and Is Regulated Developmentally and by Osmotic Signals. Plant Cell. 15(4):981-991. https://doi.org/10.1105/tpc.009787

2.5  Spacers

spacers用於在多肽和貨物之間創建一個距離，以減少多肽結合位點的立体阻礙。在這種情況下，貨物可以是藥物、染料、標籤。

2.5.1  PEGylation

將聚（乙二醇）附著到肽上稱為 PEGylation。短的雙官能 PEG（聚乙二醇）可用作肽與其他分子進行生物共結時的間隔物。PEG 生物共結技術也被用來改善多肽的蛋白水解穩定性、生物分布和溶解度^1-2。

Molecular Weight	146 g/mol
Availability：	PEPscreen®

參考資料

1. Sullivan TP, van Poll ML, Dankers PYW, Huck WTS. 2004. Forced Peptide Synthesis in Nanoscale Confinement under Elastomeric Stamps. Angew. Chem.. 116(32):4286-4289. https://doi.org/10.1002/ange.200460271

2. Veronese FM. 2001. Peptide and protein PEGylation. Biomaterials. 22(5):405-417. https://doi.org/10.1016/s0142-9612(00)00193-9

2．2  氨基己酸

氨基己酸是一種疏水墊片，可將分子（螢光體、標籤或任何生物分子）附在肽上 ^1-2.

。

Molecular Weight	113 g/mol
Availability：	PEPscreen®

參考資料

1. Mitchell D, Steinman L, Kim D, Fathman C, Rothbard J. 2000. Polyarginine enters cells more efficiently than other polycationic homopolymers. J Pept Res. 56(5):318-325. https://doi.org/10.1034/j.1399-3011.2000.00723.x

3.0 C 端修飾

3.1  酰胺（酰胺化）

肽的 C 端合成為酰胺，以中和 C 端 COOH 產生的負電荷。添加此修飾是為了防止酵素降解、模仿原生蛋白質，以及在某些情況下去除可能干擾檢測的縮氨酸 C 終端的氫鍵。

參考資料

1. Kim K, Seong BL. 2001. Peptide amidation: Production of peptide hormonesin vivo andin vitro. Biotechnol. Bioprocess Eng.. 6(4):244-251. https://doi.org/10.1007/bf02931985