.png){kind=icon}
Return
Quantifying membrane binding and diffusion with fluorescence correlation spectroscopy diffusion laws
.png){kind=icon}
OA
AI
DOI：10.1016/j.bpj.2023.01.006.png)
.png){kind=icon}
.png){kind=icon}
Share.png){kind=icon}
Save
Abstract
En 中文
Many transient processes in cells arise from the binding of cytosolic proteins to membranes. Quantifying this membrane binding and its associated diffusion in the living cell is therefore of primary importance. Dynamic photonic micros-copies, e.g., single/multiple particle tracking, fluorescence recovery after photobleaching, and fluorescence correlation spectros-copy (FCS), enable non-invasive measurement of molecular mobility in living cells and their plasma membranes. However, FCS with a single beam waist is of limited applicability with complex, non-Brownian, motions. Recently, the development of FCS diffu-sion laws methods has given access to the characterization of these complex motions, although none of them is applicable to the membrane binding case at the moment. In this study, we combined computer simulations and FCS experiments to propose an FCS diffusion law for membrane binding. First, we generated computer simulations of spot-variation FCS (svFCS) measure-ments for a membrane binding process combined to 2D and 3D diffusion at the membrane and in the bulk/cytosol, respectively. Then, using these simulations as a learning set, we derived an empirical diffusion law with three free parameters: the apparent binding constant KD, the diffusion coefficient on the membrane D2D, and the diffusion coefficient in the cytosol, D3D. Finally, we monitored, using svFCS, the dynamics of retroviral Gag proteins and associated mutants during their binding to supported lipid bilayers of different lipid composition or at plasma membranes of living cells, and we quantified KD and D2D in these conditions using our empirical diffusion law. Based on these experiments and numerical simulations, we conclude that this new approach enables correct estimation of membrane partitioning and membrane diffusion properties (KD and D2D) for peripheral membrane molecules.
Keywords:
PLASMA-MEMBRANE
HIV-1
PHOSPHATIDYLINOSITOL-(4,5)-BISPHOSPHATE
MICRODOMAINS
PROTEINS
DOMAIN
LIPIDS
MODEL
AI Summary
Key information extracted from the uploaded paper, including a brief overview, abstract, background, key highlights, visual analysis, and future outlook.
Journal
IF:
3.1
Papers:
5.1W
Citations:
4.4W
Organization
Cited Papers
ShareSaveDark states in monomeric red fluorescent proteins studied by fluorescence correlation and single molecule spectroscopy.png)
BIOPHYSICAL JOURNALOAAI
BIOPHYSICAL JOURNALIF3.1
OAAIShareSaveDiffusion analysis within single nanometric apertures reveals the ultrafine cell membrane organizationBIOPHYSICAL JOURNALOAAI
BIOPHYSICAL JOURNALIF3.1
OAAIShareSaveInfluence of progesterone and oestradiol-17 on blastocysts of the tammar wallaby (Macropus eugenii) during seasonal diapauseReproductionOAAI
ReproductionIF0
OAAIShareSaveShareSaveCharacterization of Cytoplasmic Gag-Gag Interactions by Dual-Color Z-Scan Fluorescence Fluctuation SpectroscopyBIOPHYSICAL JOURNALOAAI
BIOPHYSICAL JOURNALIF3.1
OAAIShareSaveTwo-dimensional echocardiographic determination of ventricular volumes in the fetal heart. Validation studies in fetal lambs.CirculationOAAI
CirculationIF0
OAAIShareSaveShareSave
View more