PMID- 20511226
OWN - NLM
STAT- MEDLINE
DCOM- 20100914
LR  - 20211020
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 285
IP  - 31
DP  - 2010 Jul 30
TI  - Targeting of tumor necrosis factor receptor 1 to low density plasma membrane 
      domains in human endothelial cells.
PG  - 23868-79
LID - 10.1074/jbc.M110.122853 [doi]
AB  - TNFR1 (tumor necrosis factor receptor 1) localizes to caveolae of human 
      endothelial-derived EA.hy926 cells. Transduced TNFR1 molecules lacking amino acid 
      residues 229-244 (spanning the transmembrane/intercellular boundary) are 
      expressed on the cell surface equivalently to full-length TNFR1 molecules but 
      incompletely localize to caveolae. A peptide containing this sequence pulls down 
      CAV-1 (caveolin-1) and TNFR1 from cell lysates but fails to do so following 
      disruption of caveolae with methyl-beta-cyclodextrin. We previously reported that 
      methyl-beta-cyclodextrin eliminates caveolae and blocks tumor necrosis factor 
      (TNF)-induced internalization of TNFR1 but not TNF-induced activation of 
      NF-kappaB in EA.hy926 cells. Both CAV-1 and FLOT-2 (flotillin-2), organizing 
      proteins of caveolae and lipid rafts, respectively, associate with caveolae in 
      EA.hy926 cells. Small interfering RNA-mediated knockdown of CAV-1 but not FLOT-2 
      strikingly reduces caveolae number. Both knockdowns reduce total TNFR1 protein 
      expression, but neither prevents TNFR1 localization to low density membrane 
      domains, TNF-induced internalization of TNFR1, or NF-kappaB activation by TNF. 
      Both CAV-1 and FLOT-2 knockdowns reduce TNF-mediated activation of 
      stress-activated protein kinase (SAPK). However, both knockdowns reduce 
      expression of TRAF2 (TNF receptor-associated factor-2) protein, and small 
      interfering RNA targeting of TRAF2 also selectively inhibits SAPK activation. We 
      conclude that TNFR1 contains a membrane-proximal sequence that targets the 
      receptor to caveolae/lipid rafts. Neither TNFR1 targeting to nor internalization 
      from these low density membrane domains depends upon CAV-1 or FLOT-2. 
      Furthermore, both NF-kappaB and SAPK activation appear independent of both TNFR1 
      localization to low density membrane domains and to TNF-induced receptor 
      internalization.
FAU - D'Alessio, Alessio
AU  - D'Alessio A
AD  - Interdepartmental Program in Vascular Biology and Therapeutics, Yale University 
      School of Medicine, New Haven, CT 06520-8089, USA.
FAU - Kluger, Martin S
AU  - Kluger MS
FAU - Li, Jie H
AU  - Li JH
FAU - Al-Lamki, Rafia
AU  - Al-Lamki R
FAU - Bradley, John R
AU  - Bradley JR
FAU - Pober, Jordan S
AU  - Pober JS
LA  - eng
GR  - BHF_/British Heart Foundation/United Kingdom
GR  - N01HV28186/HL/NHLBI NIH HHS/United States
GR  - R01 HL036003/HL/NHLBI NIH HHS/United States
GR  - N01-HV28186/HV/NHLBI NIH HHS/United States
GR  - R01-HL36003/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20100528
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (NF-kappa B)
RN  - 0 (Peptides)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (Receptors, Tumor Necrosis Factor, Type I)
RN  - 0 (beta-Cyclodextrins)
RN  - 0 (methyl-beta-cyclodextrin)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinase 8)
SB  - IM
MH  - Caveolae/metabolism
MH  - Cell Membrane/*metabolism
MH  - Cell Separation
MH  - Endothelial Cells/*cytology
MH  - Flow Cytometry
MH  - Fluorescent Antibody Technique, Indirect
MH  - Humans
MH  - Membrane Microdomains/chemistry
MH  - Mitogen-Activated Protein Kinase 8/metabolism
MH  - NF-kappa B/metabolism
MH  - Peptides/chemistry
MH  - RNA, Small Interfering/metabolism
MH  - Receptors, Tumor Necrosis Factor, Type I/*metabolism
MH  - Signal Transduction
MH  - beta-Cyclodextrins/chemistry
PMC - PMC2911292
EDAT- 2010/06/01 06:00
MHDA- 2010/09/16 06:00
PMCR- 2011/07/30
CRDT- 2010/06/01 06:00
PHST- 2010/06/01 06:00 [entrez]
PHST- 2010/06/01 06:00 [pubmed]
PHST- 2010/09/16 06:00 [medline]
PHST- 2011/07/30 00:00 [pmc-release]
AID - S0021-9258(20)61853-1 [pii]
AID - M110.122853 [pii]
AID - 10.1074/jbc.M110.122853 [doi]
PST - ppublish
SO  - J Biol Chem. 2010 Jul 30;285(31):23868-79. doi: 10.1074/jbc.M110.122853. Epub 
      2010 May 28.