Detail of Fig2A_pten3



Project
Title
Time-lapse TIRF microscopy images of D. discoideum cells expressing PTEN mutants labeled with TMR
Description
NA
Release, Updated
2017-10-03,
2018-11-15
License
CC BY
Kind
Image data based on Experiment
File Formats
Data size
150.0 MB

Organism
D. discoideum ( NCBI:txid44689 )
Strain(s)
-
Cell Line
-
Protein names
PTEN

Datatype
single molecule dynamics
Molecular Function (MF)
Biological Process (BP)
chemotaxis ( GO:0006935 )
Cellular Component (CC)
-
Biological Imaging Method
XYZ Scale
XY: 0.033 micrometer/pixel, Z: NA
T scale
33 millisecond for each time interval

Image Acquisition
Experiment type
TimeLapse
Microscope type
TotalInternalReflectionFluorescenceMicroscope
Acquisition mode
TIRF
Contrast method
Fluorescence
Microscope model
OLYMPUS IX-70
Detector model
Andor iXon3
Objective model
OLYMPUS PlanApo 60x 1.45NA Oil TIRFM
Filter set

Summary of Methods
See details in Yasui et al. (2014) PLoS Computational Biology, 10(9): e1003817.
Related paper(s)

Masato Yasui, Satomi Matsuoka, Masahiro Ueda (2014) PTEN hopping on the cell membrane is regulated via a positively-charged C2 domain., PLoS computational biology, Volume 10, Number 9, pp. e1003817

Published in 2014 Sep (Electronic publication in Sept. 11, 2014, midnight )

(Abstract) PTEN, a tumor suppressor that is frequently mutated in a wide spectrum of cancers, exerts PI(3,4,5)P3 phosphatase activities that are regulated by its dynamic shuttling between the membrane and cytoplasm. Direct observation of PTEN in the interfacial environment can offer quantitative information about the shuttling dynamics, but remains elusive. Here we show that positively charged residues located in the calpha2 helix of the C2 domain are necessary for the membrane localization of PTEN via stable electrostatic interactions in Dictyostelium discoideum. Single-molecule imaging analyses revealed that PTEN molecules moved distances much larger than expected had they been caused by lateral diffusion, a phenomenon we call "hopping." Our novel single-particle tracking analysis method found that the calpha2 helix aids in regulating the hopping and stable-binding states. The dynamically established membrane localization of PTEN was revealed to be essential for developmental processes and clarified a fundamental regulation mechanism of the protein quantity and activity on the plasma membrane.
(MeSH Terms)

Contact
Satomi Matsuoka , Osaka University , Graduate School of Frontier Biosciences , Laboratories for Nanobiology
Contributors
Masato Yasui, Satomi Matsuoka, Masahiro Ueda

OMERO Dataset
OMERO Project
Source