PMID- 15860728
OWN - NLM
STAT- MEDLINE
DCOM- 20050919
LR  - 20111117
IS  - 0021-9533 (Print)
IS  - 0021-9533 (Linking)
VI  - 118
IP  - Pt 9
DP  - 2005 May 1
TI  - Single-molecule diffusion measurements of H-Ras at the plasma membrane of live 
      cells reveal microdomain localization upon activation.
PG  - 1799-809
AB  - Recent studies show that the partitioning of the small GTPase H-Ras in different 
      types of membrane microdomains is dependent on guanosine 5'-triphosphate 
      (GTP)-loading of H-Ras. Detailed knowledge about the in vivo dynamics of this 
      phenomenon is limited. In this report, the effect of the activation of H-Ras on 
      its microdomain localization was studied by single-molecule fluorescence 
      microscopy. Individual human H-Ras molecules fused to the enhanced yellow 
      fluorescent protein (eYFP) were imaged in the dorsal plasma membrane of live 
      mouse cells and their diffusion behavior was analyzed. The diffusion of a 
      constitutively inactive (S17N) and constitutively active (G12V) mutant of H-Ras 
      was compared. Detailed analysis revealed that for both mutants a major, 
      fast-diffusing population and a minor, slow-diffusing population were present. 
      The slow-diffusing fraction of the active mutant was confined to 200 nm domains, 
      which were not observed for the inactive mutant. In line with these results we 
      observed that the slow-diffusing fraction of wild-type H-Ras became confined to 
      200 nm domains upon insulin-induced activation of wild-type H-Ras. This 
      activation-dependent localization of H-Ras to 200 nm domains, for the first time 
      directly detected in live cells, supports the proposed relationship between H-Ras 
      microdomain localization and activation.
FAU - Lommerse, Piet H M
AU  - Lommerse PH
AD  - Department of Biophysics, Leiden Institute of Physics, Leiden University, Niels 
      Bohrweg 2, 2333 CA Leiden, The Netherlands.
FAU - Snaar-Jagalska, B Ewa
AU  - Snaar-Jagalska BE
FAU - Spaink, Herman P
AU  - Spaink HP
FAU - Schmidt, Thomas
AU  - Schmidt T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Cell Sci
JT  - Journal of cell science
JID - 0052457
RN  - 0 (Bacterial Proteins)
RN  - 0 (Insulin)
RN  - 0 (Luminescent Proteins)
RN  - 0 (yellow fluorescent protein, Bacteria)
RN  - 86-01-1 (Guanosine Triphosphate)
RN  - EC 3.6.5.2 (ras Proteins)
SB  - IM
MH  - Animals
MH  - Bacterial Proteins/metabolism
MH  - Biophysics/methods
MH  - Cell Line
MH  - Cell Membrane/*metabolism
MH  - Cytoskeleton/metabolism
MH  - Diffusion
MH  - Fibroblasts/metabolism
MH  - Fluorescence Resonance Energy Transfer
MH  - Genes, ras/*genetics
MH  - Guanosine Triphosphate/chemistry/metabolism
MH  - Humans
MH  - Insulin/metabolism
MH  - Luminescent Proteins/metabolism
MH  - MAP Kinase Signaling System
MH  - Mice
MH  - Microscopy, Fluorescence/*methods
MH  - Models, Statistical
MH  - Mutation
MH  - Plasmids/metabolism
MH  - Protein Structure, Tertiary
MH  - Time Factors
MH  - Transfection
MH  - ras Proteins/*chemistry/metabolism
EDAT- 2005/04/30 09:00
MHDA- 2005/09/20 09:00
CRDT- 2005/04/30 09:00
PHST- 2005/04/30 09:00 [pubmed]
PHST- 2005/09/20 09:00 [medline]
PHST- 2005/04/30 09:00 [entrez]
AID - 118/9/1799 [pii]
AID - 10.1242/jcs.02300 [doi]
PST - ppublish
SO  - J Cell Sci. 2005 May 1;118(Pt 9):1799-809. doi: 10.1242/jcs.02300.