Multiplex detection of histone-modifying enzymes by total internal reflection fluorescence-based single-molecule detection

被引:49
|
作者
Ma, Fei [1 ,2 ]
Liu, Meng [1 ]
Wang, Zi-yue [1 ]
Zhang, Chun-yang [1 ,2 ]
机构
[1] Shandong Normal Univ, Coll Chem Chem Engn & Mat Sci, Collaborat Innovat Ctr Functionalized Probes Chem, Key Lab Mol & Nano Probes,Minist Educ,Shandong Pr, Jinan 250014, Peoples R China
[2] Chinese Acad Sci, Shenzhen Inst Adv Technol, Single Mol Detect & Imaging Lab, Shenzhen 518055, Peoples R China
基金
中国国家自然科学基金;
关键词
PROTEIN-PHOSPHORYLATION; DNA-REPLICATION; ASSAY; ACETYLATION; METHYLTRANSFERASES; GCN5; H3; ACETYLTRANSFERASES; SPECIFICITY; COMPLEXES;
D O I
10.1039/c5cc08797j
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
We develop a sensitive and selective method for the multiplex detection of histone-modifying enzymes (HMEs) through the integration of antibody-based fluorescence labeling with total internal reflection fluorescence (TIRF)-based single-molecule detection. This method exhibits excellent specificity and high sensitivity with a detection limit of 21 pM for histone acetyltransferase GcN5 and 12 pM for histone methyltransferase G9a, and it can be applied for the screening of HME inhibitors as well.
引用
收藏
页码:1218 / 1221
页数:4
相关论文
共 50 条
  • [31] Detection efficiency in total internal reflection fluorescence microscopy
    Leutenegger, Marcel
    Lasser, Theo
    OPTICS EXPRESS, 2008, 16 (12) : 8519 - 8531
  • [32] Single-molecule detection and identification of multiple species by multiparameter fluorescence detection
    Widengren, J
    Kudryavtsev, V
    Antonik, M
    Berger, S
    Gerken, M
    Seidel, CAM
    ANALYTICAL CHEMISTRY, 2006, 78 (06) : 2039 - 2050
  • [33] Multiplex detection of lung cancer cells at the single-molecule level
    Hu, Juan
    Zhang, Chun-yang
    CHEMICAL COMMUNICATIONS, 2014, 50 (88) : 13581 - 13584
  • [34] Enhancement of single-molecule fluorescence detection in subwavelength apertures
    Rigneault, H
    Capoulade, J
    Dintinger, J
    Wenger, J
    Bonod, N
    Popov, E
    Ebbesen, TW
    Lenne, PF
    PHYSICAL REVIEW LETTERS, 2005, 95 (11)
  • [35] Single-molecule fluorescence detection of a tricyclic nucleoside analogue
    Samaan, George N.
    Wyllie, Mckenzie K.
    Cizmic, Julian M.
    Needham, Lisa-Maria
    Nobis, David
    Ngo, Katrina
    Andersen, Susan
    Magennis, Steven W.
    Lee, Steven F.
    Purse, Byron W.
    CHEMICAL SCIENCE, 2021, 12 (07) : 2623 - 2628
  • [36] A Starting Point for Fluorescence-Based Single-Molecule Measurements in Biomolecular Research
    Gust, Alexander
    Zander, Adrian
    Gietl, Andreas
    Holzmeister, Phil
    Schulz, Sarah
    Lalkens, Birka
    Tinnefeld, Philip
    Grohmann, Dina
    MOLECULES, 2014, 19 (10): : 15824 - 15865
  • [37] Subcellular and single-molecule imaging of plant fluorescent proteins using total internal reflection fluorescence microscopy (TIRFM)
    Vizcay-Barrena, Gema
    Webb, Stephen E. D.
    Martin-Fernandez, Marisa L.
    Wilson, Zoe A.
    JOURNAL OF EXPERIMENTAL BOTANY, 2011, 62 (15) : 5419 - 5428
  • [38] Trends in Single-Molecule Total Internal Reflection Fluorescence Imaging and Their Biological Applications with Lab-on-a-Chip Technology
    Colson, Louis
    Kwon, Youngeun
    Nam, Soobin
    Bhandari, Avinashi
    Maya, Nolberto Martinez
    Lu, Ying
    Cho, Yongmin
    SENSORS, 2023, 23 (18)
  • [39] A comparison of through-the-objective total internal reflection microscopy and epifluorescence microscopy for single-molecule fluorescence Imaging
    Paige, MF
    Bjerneld, EJ
    Moerner, WE
    SINGLE MOLECULES, 2001, 2 (03) : 191 - 201
  • [40] Fluorescence-Based Multiplex Protein Detection Using Optically Encoded Microbeads
    Jun, Bong-Hyun
    Kang, Homan
    Lee, Yoon-Sik
    Jeong, Dae Hong
    MOLECULES, 2012, 17 (03) : 2474 - 2490