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
来源：
CHEMICAL COMMUNICATIONS | 2016年 / 52卷 / 06期
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
  • [21] Illuminating amyloid fibrils: Fluorescence-based single-molecule approaches
    Rice, Lauren J.
    Ecroyd, Heath
    van Oijen, Antoine M.
    COMPUTATIONAL AND STRUCTURAL BIOTECHNOLOGY JOURNAL, 2021, 19 : 4711 - 4724
  • [22] Cool and dynamic: single-molecule fluorescence-based structural biology
    Craggs, Timothy D.
    NATURE METHODS, 2017, 14 (02) : 123 - 124
  • [23] Single Molecule Detection at Surfaces: Dual-Color Fluorescence Fluctuation Spectroscopy with Total Internal Reflection Excitation
    Leutenegger, M.
    Hassler, K.
    Rigler, P.
    Bilenca, A.
    Lasser, T.
    NEW APPROACHES IN BIOMEDICAL SPECTROSCOPY, 2007, 963 : 259 - 279
  • [24] Cool and dynamic: single-molecule fluorescence-based structural biology
    Timothy D Craggs
    Nature Methods, 2017, 14 : 123 - 124
  • [25] Total internal reflection fluorescence microscopy in single molecule nanobioscience
    Wazawa, T
    Ueda, M
    MICROSCOPY TECHNIQUES, 2005, 95 : 77 - 106
  • [26] Recent advance in optical single-molecule detection of methylation modification and methyl-modifying enzymes
    Han, Yun
    Yang, Dongming
    Jiang, Su
    Zhao, Shulin
    Ma, Fei
    Zhang, Chun-yang
    TRAC-TRENDS IN ANALYTICAL CHEMISTRY, 2024, 172
  • [27] Total internal reflection-based module for fluorescence and absorbance detection
    Verschooten, Tom
    Ottevaere, Heidi
    Vervaeke, Michael
    Van Erps, Juergen
    Thienpont, Hugo
    JOURNAL OF MICRO-NANOLITHOGRAPHY MEMS AND MOEMS, 2014, 13 (03):
  • [28] Multidimensional single-molecule imaging in live cells using total-internal-reflection fluorescence microscopy
    Webb, S. E. D.
    Needham, S. R.
    Roberts, S. K.
    Martin-Fernandez, M. L.
    OPTICS LETTERS, 2006, 31 (14) : 2157 - 2159
  • [29] Single-Molecule Study of Actin Filament Severing by Gelsolin using Total Internal Reflection Fluorescence Microscopy
    Kannan, Balakrishnan
    Lee, Wei Lin
    Robinson, Robert C.
    BIOPHYSICAL JOURNAL, 2010, 98 (03) : 155A - 155A
  • [30] Modules for teaching and applying single-molecule total internal reflection fluorescence microscopy in an undergraduate lab course
    Knight, Jefferson
    Alansari, Noha
    Giardina, Daniel T.
    Huynh, Thao N.
    Saunders, Emma
    BIOPHYSICAL JOURNAL, 2024, 123 (03) : 308A - 308A
12345