Evaluation of diffusion coefficient of P-glycoprotein molecules labeled with green fluorescent protein in living cell membrane.
| Title: | Evaluation of diffusion coefficient of P-glycoprotein molecules labeled with green fluorescent protein in living cell membrane. |
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| Authors: | Vu XH; Institute of Science and Technology, TNU- University of Sciences (TNUS), Tan Thinh ward, Thai Nguyen city, Viet Nam.; Dien ND; Faculty of Labour Protection, Vietnam Trade Union University, 169 Tay Son street, Hanoi city, Viet Nam.; Pham TTH; Faculty of Chemistry, TNU- University of Sciences (TNUS), Tan Thinh ward, Thai Nguyen city, Viet Nam. Electronic address: haptt@tnus.edu.vn.; Jaffiol R; Laboratoire de Nanotechnologie et d'Instrumentation Optique, Institut Charles Delaunay, UMR CNRS 6281, Université de Technologie de Troyes, 12 Rue Marie Curie, CS 42060, 10 004 Troyes Cedex, France. Electronic address: rodolphe.jaffiol@utt.fr.; Vézy C; Laboratoire de Nanotechnologie et d'Instrumentation Optique, Institut Charles Delaunay, UMR CNRS 6281, Université de Technologie de Troyes, 12 Rue Marie Curie, CS 42060, 10 004 Troyes Cedex, France.; Ca NX; Institute of Science and Technology, TNU- University of Sciences (TNUS), Tan Thinh ward, Thai Nguyen city, Viet Nam.; Trang TT; Institute of Science and Technology, TNU- University of Sciences (TNUS), Tan Thinh ward, Thai Nguyen city, Viet Nam. |
| Source: | Biochimica et biophysica acta. Biomembranes [Biochim Biophys Acta Biomembr] 2021 Nov 01; Vol. 1863 (11), pp. 183721. Date of Electronic Publication: 2021 Aug 02. |
| Publication Type: | Journal Article; Research Support, Non-U.S. Gov't |
| Language: | English |
| Journal Info: | Publisher: Elsevier Country of Publication: Netherlands NLM ID: 101731713 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-2642 (Electronic) Linking ISSN: 00052736 NLM ISO Abbreviation: Biochim Biophys Acta Biomembr Subsets: MEDLINE |
| Imprint Name(s): | Original Publication: Amsterdam : Elsevier |
| MeSH Terms: | ATP Binding Cassette Transporter, Subfamily B, Member 1/*metabolism ; Green Fluorescent Proteins/*metabolism; Cell Membrane/metabolism ; Microscopy, Fluorescence/methods ; Animals ; Diffusion ; Dogs ; Madin Darby Canine Kidney Cells |
| Abstract: | The movement of individual molecules inside living cells has recently been resolved by single particles tracking (SPT) method which is a powerful tool for probing the organization and dynamics of the plasma membrane constituents. Effective treatment of metastatic cancers requires the toxic chemotherapy, however this therapy leads to the multidrug resistance phenomenon of the cancer cells, in which the cancer cells resist simultaneously to different drugs with different targets and chemical structures. P-glycoprotein molecules which are responsible for multidrug resistance of many cancer cells have been studied by cancer biologists during past haft of century. Recently, advances in laser and detector technologies have enabled single fluorophores to be visualized in aqueous solution. The development of the total internal reflection fluorescent microscope (TIRFM) provided means to monitor dynamic molecular localization in living cells. In this paper, P-glycoproteins (PGP) were labeled with green fluorescent protein (GFP) in living cell membrane of Madin-Darby canine kidney (MDCK) and the TIRFM method was used to characterize the dynamics of individual protein molecules on the surface of living cells. An evanescent field was produced by a totally internally reflected and a laser beam was illuminated the glass-water interface. GFP-PGP proteins that entered the evanescent field appeared as individual spots of light which were slighter than background fluorescence. We obtained high-resolution images and diffusion maps of membrane proteins on cell surface and showed the local diffusion properties of specific proteins on single cells. We also determined the diffusion coefficient, the mean square displacement and the average velocity of the tracked particles, as well as the heterogeneity of the cell environment. This study enabled us to understand single-molecule features in living cell and measure the diffusion kinetics of membrane-bound molecules.; (Copyright © 2021 Elsevier B.V. All rights reserved.) |
| Contributed Indexing: | Keywords: Green fluorescent protein (GFP); Living cell membrane; P-glycoprotein (PGP); Single-particle tracking (SPT); Total internal reflection fluorescent microscopy (TIRFM) |
| Substance Nomenclature: | 0 (ATP Binding Cassette Transporter, Subfamily B, Member 1); 147336-22-9 (Green Fluorescent Proteins) |
| Entry Date(s): | Date Created: 20210805 Date Completed: 20211117 Latest Revision: 20211117 |
| Update Code: | 20260130 |
| DOI: | 10.1016/j.bbamem.2021.183721 |
| PMID: | 34352241 |
| Database: | MEDLINE |
Journal Article; Research Support, Non-U.S. Gov't