Probing SH2-domains using Inhibitor Affinity Purification (IAP)
نویسندگان
چکیده
منابع مشابه
High-throughput phosphotyrosine profiling using SH2 domains.
Protein tyrosine phosphorylation controls many aspects of signaling in multicellular organisms. One of the major consequences of tyrosine phosphorylation is the creation of binding sites for proteins containing Src homology 2 (SH2) domains. To profile the global tyrosine phosphorylation state of the cell, we have developed proteomic binding assays encompassing nearly the full complement of huma...
متن کاملProtein-phosphotyrosine proteome profiling by superbinder-SH2 domain affinity purification mass spectrometry, sSH2-AP-MS.
Recently, "superbinder" SH2 domain variants with three amino acid substitutions (sSH2) were reported to have 100-fold or greater affinity for protein-phosphotyrosine (pY) than natural SH2 domains. Here we report a protocol in which His-tagged Src sSH2 efficiently captures pY-peptides from protease-digested HeLa cell total protein extracts. Affinity purification of pY-peptides by this method sho...
متن کاملProbing the phosphopeptide specificities of protein tyrosine phosphatases, SH2 and PTB domains with combinatorial library methods.
Protein tyrosine phosphatases, SH2 and PTB domains are crucial elements for cellular signal transduction and regulation. Much effort has been directed towards elucidating their specificity in the past decade using a variety of approaches. Combinatorial library methods have contributed significantly to the understanding of substrate and ligand specificity of phosphoprotein recognizing domains. T...
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Stereomicroscope The new Carl Zeiss stereomicroscope Stemi 2000-CS allows the user to view the sample whilst other observers view the procedure by video. Greenough type stereomicroscopes create their stereo effect by having separate light paths set at different angles, one for each eye. However, when video or photomicrography is being carried out most stereomicroscope systems remove light from ...
متن کاملProtein purification using PDZ affinity chromatography.
PDZ domains function in nature as protein-binding domains within scaffold and membrane-associated proteins. They comprise approximately 90 residues and undergo specific, high-affinity interactions with complementary C-terminal peptide sequences, other PDZ domains, and/or phospholipids. We have previously shown that the specific, strong interactions of PDZ domains with their ligands make them we...
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ژورنال
عنوان ژورنال: Proteome Science
سال: 2014
ISSN: 1477-5956
DOI: 10.1186/1477-5956-12-41