PMID- 10085250
OWN - NLM
STAT- MEDLINE
DCOM- 19990719
LR  - 20220215
IS  - 0021-9533 (Print)
IS  - 0021-9533 (Linking)
VI  - 112 ( Pt 8)
DP  - 1999 Apr
TI  - Immunofluorescence detection of ezrin/radixin/moesin (ERM) proteins with their 
      carboxyl-terminal threonine phosphorylated in cultured cells and tissues.
PG  - 1149-58
AB  - Ezrin/radixin/moesin (ERM) proteins are thought to play an important role in 
      organizing cortical actin-based cytoskeletons through cross-linkage of actin 
      filaments with integral membrane proteins. Recent in vitro biochemical studies 
      have revealed that ERM proteins phosphorylated on their COOH-terminal threonine 
      residue (CPERMs) are active in their cross-linking activity, but this has not yet 
      been evaluated in vivo. To immunofluorescently visualize CPERMs in cultured cells 
      as well as tissues using a mAb specific for CPERMs, we developed a new fixation 
      protocol using trichloroacetic acid (TCA) as a fixative. Immunoblotting analyses 
      in combination with immunofluorescence microscopy showed that TCA effectively 
      inactivated soluble phosphatases, which maintained the phosphorylation level of 
      CPERMs during sample processing for immunofluorescence staining. 
      Immunofluorescence microscopy with TCA-fixed samples revealed that CPERMs were 
      exclusively associated with plasma membranes in a variety of cells and tissues, 
      whereas total ERM proteins were distributed in both the cytoplasm and plasma 
      membranes. Furthermore, the amounts of CPERMs were shown to be regulated in a 
      cell and tissue type-dependent manner. These findings favored the notion that 
      phosphorylation of the COOH-terminal threonine plays a key role in the regulation 
      of the cross-linking activity of ERM proteins in vivo.
FAU - Hayashi, K
AU  - Hayashi K
AD  - Department of Cell Biology, Faculty of Medicine, Kyoto University, Sakyo-ku, 
      Kyoto 606-8501, Japan.
FAU - Yonemura, S
AU  - Yonemura S
FAU - Matsui, T
AU  - Matsui T
FAU - Tsukita, S
AU  - Tsukita S
LA  - eng
PT  - Comparative Study
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 (Blood Proteins)
RN  - 0 (Cytoskeletal Proteins)
RN  - 0 (Membrane Proteins)
RN  - 0 (Microfilament Proteins)
RN  - 0 (Phosphoproteins)
RN  - 0 (ezrin)
RN  - 1364PS73AF (Acetone)
RN  - 144131-77-1 (moesin)
RN  - 144517-21-5 (radixin)
RN  - 1HG84L3525 (Formaldehyde)
RN  - 3K9958V90M (Ethanol)
RN  - 5V2JDO056X (Trichloroacetic Acid)
RN  - Y4S76JWI15 (Methanol)
SB  - IM
MH  - 3T3 Cells
MH  - Acetone/metabolism
MH  - Animals
MH  - Blood Proteins/*analysis/immunology
MH  - Brain/metabolism
MH  - Cells, Cultured
MH  - *Cytoskeletal Proteins
MH  - Dogs
MH  - Epithelial Cells/metabolism
MH  - Ethanol/metabolism
MH  - Female
MH  - Formaldehyde/metabolism
MH  - Humans
MH  - Hypoxia
MH  - Immunoblotting
MH  - Intestine, Small/metabolism
MH  - Kidney/metabolism
MH  - Liver/metabolism
MH  - Membrane Proteins/*analysis/immunology
MH  - Methanol/metabolism
MH  - Mice
MH  - Microfilament Proteins/*analysis/immunology
MH  - Microscopy, Confocal
MH  - Microscopy, Fluorescence/*methods
MH  - Ovary/metabolism
MH  - Phosphoproteins/*analysis/immunology
MH  - Phosphorylation
MH  - Rats
MH  - Sciatic Nerve/metabolism
MH  - Tongue/metabolism
MH  - Trichloroacetic Acid/*chemistry
MH  - Tumor Cells, Cultured
EDAT- 1999/03/23 00:00
MHDA- 1999/03/23 00:01
CRDT- 1999/03/23 00:00
PHST- 1999/03/23 00:00 [pubmed]
PHST- 1999/03/23 00:01 [medline]
PHST- 1999/03/23 00:00 [entrez]
AID - 10.1242/jcs.112.8.1149 [doi]
PST - ppublish
SO  - J Cell Sci. 1999 Apr;112 ( Pt 8):1149-58. doi: 10.1242/jcs.112.8.1149.