Source:http://linkedlifedata.com/resource/pubmed/id/19754663
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
11
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| pubmed:dateCreated |
2010-11-30
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| pubmed:abstractText |
?-Secretase is a transmembrane protease complex responsible for the processing of a multitude of type 1 transmembrane proteins, including amyloid precursor protein (APP) and Notch. A functional complex is dependent on the assembly of four proteins: presenilin (PS), nicastrin, Aph-1 and Pen-2. Little is known about how the substrates are selected by ?-secretase, but it has been suggested that ?-secretase associated proteins (GSAPs) could be of importance. For instance, it was recently reported from studies in cell lines that TMP21, a transmembrane protein involved in trafficking, binds to ?-secretase and regulates the processing of APP-derived substrates without affecting Notch cleavage. Here, we present an efficient and selective method for purification and analysis of ?-secretase and GSAPs. Microsomal membranes were prepared from rat or human brain and incubated with a ?-secretase inhibitor coupled to biotin via a long linker and a S-S bridge. After pulldown using streptavidin beads, bound proteins were eluted under reducing conditions and digested by trypsin. The tryptic peptides were subjected to LC-MS/MS analysis, and proteins were identified by sequence data from MS/MS spectra. All of the known ?-secretase components were identified. Interestingly, TMP21 and the PS associated protein syntaxin1 were associated to ?-secretase in rat brain. We suggest that the present method can be used for further studies on the composition of the ?-secretase complex.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Amyloid Precursor Protein Secretases,
http://linkedlifedata.com/resource/pubmed/chemical/Enzyme Inhibitors,
http://linkedlifedata.com/resource/pubmed/chemical/Membrane Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Peptide Fragments,
http://linkedlifedata.com/resource/pubmed/chemical/Presenilin-1,
http://linkedlifedata.com/resource/pubmed/chemical/Syntaxin 1,
http://linkedlifedata.com/resource/pubmed/chemical/TMP21 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Tmp21 protein, rat
|
| pubmed:status |
MEDLINE
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| pubmed:month |
Nov
|
| pubmed:issn |
1582-4934
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| pubmed:author | |
| pubmed:copyrightInfo |
© 2009 The Authors Journal compilation © 2010 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd.
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| pubmed:issnType |
Electronic
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| pubmed:volume |
14
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
2675-86
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| pubmed:meshHeading |
pubmed-meshheading:19754663-Amino Acid Sequence,
pubmed-meshheading:19754663-Amyloid Precursor Protein Secretases,
pubmed-meshheading:19754663-Animals,
pubmed-meshheading:19754663-Blotting, Western,
pubmed-meshheading:19754663-Brain,
pubmed-meshheading:19754663-Chromatography, Affinity,
pubmed-meshheading:19754663-Chromatography, Liquid,
pubmed-meshheading:19754663-Enzyme Inhibitors,
pubmed-meshheading:19754663-Humans,
pubmed-meshheading:19754663-Membrane Proteins,
pubmed-meshheading:19754663-Microsomes,
pubmed-meshheading:19754663-Molecular Sequence Data,
pubmed-meshheading:19754663-Peptide Fragments,
pubmed-meshheading:19754663-Presenilin-1,
pubmed-meshheading:19754663-Rats,
pubmed-meshheading:19754663-Rats, Sprague-Dawley,
pubmed-meshheading:19754663-Syntaxin 1,
pubmed-meshheading:19754663-Tandem Mass Spectrometry
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| pubmed:year |
2010
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| pubmed:articleTitle |
Affinity pulldown of ?-secretase and associated proteins from human and rat brain.
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| pubmed:affiliation |
The Karolinska Institutet (KI) Dainippon Sumitomo Pharma Alzheimer Center (KASPAC), KI-Alzheimer's Disease Research Center, Department of Neurobiology, Care Sciences and Society, Karolinska Institutet, Novum, Huddinge, Sweden.
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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