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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
24
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pubmed:dateCreated |
2005-6-13
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pubmed:abstractText |
The voltage-gated sodium channel beta2-subunit (beta2) is a member of the IgCAM superfamily and serves as both an adhesion molecule and an auxiliary subunit of the voltage-gated sodium channel. Here we found that beta2 undergoes ectodomain shedding followed by presenilin (PS)-dependent gamma-secretase-mediated cleavage. 12-O-Tetradecanoylphorbol-13-acetate treatment or expression of an alpha-secretase enzyme, ADAM10, resulted in ectodomain cleavage of beta2 in Chinese hamster ovary cells. Subsequent cleavage of the remaining 15-kDa C-terminal fragment (beta2-CTF) was independently inhibited by three specific gamma-secretase inhibitors, expression of the dominant negative form of PS1, and in PS1/PS2 knock-out cells. gamma-Secretase inhibitor treatment also increased endogenous beta2-CTF levels in neuroblastoma cells and mouse primary neuronal cultures. In a cell-free gamma-secretase assay, we detected gamma-secretase activity-dependent generation of a 12 kDa beta2 intracellular domain (ICD), which was loosely associated with the membrane fraction. To assess the functional role of beta2 processing by gamma-secretase, we tested whether N-[N-(3,5-difluorophenylacetyl-l-alanyl)]-S-phenylglycine t-butylester (DAPT), a specific gamma-secretase inhibitor, would alter beta2-mediated cell adhesion and migration. We found that DAPT inhibited cell-cell aggregation and migration in a wound healing assay carried out with Chinese hamster ovary cells expressing beta2. DAPT also reduced migration of neuroblastoma cells in a modified Boyden chamber assay. Since DAPT treatment resulted in increased beta2-CTF levels, we also tested whether beta2-CTFs or beta2-ICDs would directly affect cell migration by overexpressing recombinant proteins. Interestingly, elevated levels of beta2-CTFs, but not ICDs, also blocked cell migration by 81 to 93%. Together, our findings show for the first time that beta2 is a PS/gamma-secretase substrate and gamma-secretase mediated cleavage of beta2-CTF is required for cell-cell adhesion and migration of beta2-expressing cells.
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pubmed:commentsCorrections |
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/1,2-dilinolenoyl-3-(4-aminobutyryl)p...,
http://linkedlifedata.com/resource/pubmed/chemical/Amyloid Precursor Protein Secretases,
http://linkedlifedata.com/resource/pubmed/chemical/Aspartic Acid Endopeptidases,
http://linkedlifedata.com/resource/pubmed/chemical/BACE1 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Bace1 protein, mouse,
http://linkedlifedata.com/resource/pubmed/chemical/Endopeptidases,
http://linkedlifedata.com/resource/pubmed/chemical/Enzyme Inhibitors,
http://linkedlifedata.com/resource/pubmed/chemical/Membrane Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Nerve Tissue Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/PSEN1 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Presenilin-1,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Subunits,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/SCN2B protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Sodium Channels,
http://linkedlifedata.com/resource/pubmed/chemical/Tetradecanoylphorbol Acetate,
http://linkedlifedata.com/resource/pubmed/chemical/Triglycerides,
http://linkedlifedata.com/resource/pubmed/chemical/gamma-Aminobutyric Acid
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pubmed:status |
MEDLINE
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pubmed:month |
Jun
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pubmed:issn |
0021-9258
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
17
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pubmed:volume |
280
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
23251-61
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:15833746-Amino Acid Sequence,
pubmed-meshheading:15833746-Amyloid Precursor Protein Secretases,
pubmed-meshheading:15833746-Animals,
pubmed-meshheading:15833746-Aspartic Acid Endopeptidases,
pubmed-meshheading:15833746-Blotting, Western,
pubmed-meshheading:15833746-CHO Cells,
pubmed-meshheading:15833746-Cell Adhesion,
pubmed-meshheading:15833746-Cell Communication,
pubmed-meshheading:15833746-Cell Culture Techniques,
pubmed-meshheading:15833746-Cell Membrane,
pubmed-meshheading:15833746-Cell Movement,
pubmed-meshheading:15833746-Cell-Free System,
pubmed-meshheading:15833746-Cerebral Cortex,
pubmed-meshheading:15833746-Cricetinae,
pubmed-meshheading:15833746-Endopeptidases,
pubmed-meshheading:15833746-Enzyme Inhibitors,
pubmed-meshheading:15833746-Humans,
pubmed-meshheading:15833746-Membrane Proteins,
pubmed-meshheading:15833746-Mice,
pubmed-meshheading:15833746-Microscopy, Confocal,
pubmed-meshheading:15833746-Molecular Sequence Data,
pubmed-meshheading:15833746-Nerve Tissue Proteins,
pubmed-meshheading:15833746-Neurons,
pubmed-meshheading:15833746-Presenilin-1,
pubmed-meshheading:15833746-Protein Binding,
pubmed-meshheading:15833746-Protein Structure, Tertiary,
pubmed-meshheading:15833746-Protein Subunits,
pubmed-meshheading:15833746-Recombinant Proteins,
pubmed-meshheading:15833746-Sequence Homology, Amino Acid,
pubmed-meshheading:15833746-Sodium Channels,
pubmed-meshheading:15833746-Tetradecanoylphorbol Acetate,
pubmed-meshheading:15833746-Triglycerides,
pubmed-meshheading:15833746-Wound Healing,
pubmed-meshheading:15833746-gamma-Aminobutyric Acid
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pubmed:year |
2005
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pubmed:articleTitle |
Presenilin/gamma-secretase-mediated cleavage of the voltage-gated sodium channel beta2-subunit regulates cell adhesion and migration.
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pubmed:affiliation |
Neurobiology of Disease Laboratory, Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Charlestown, 02129, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't,
Research Support, N.I.H., Extramural
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