title: Preparation of recombinant protein spotted arrays for proteome-wide identification of kinase targets.
abstract: Protein microarrays allow unique approaches for interrogating global protein interaction networks. Protein arrays can be divided into two categories: antibody arrays and functional protein arrays. Antibody arrays consist of various antibodies and are appropriate for profiling protein abundance and modifications. Functional full-length protein arrays employ full-length proteins with various post-translational modifications. A key advantage of the latter is rapid parallel processing of large number of proteins for studying highly controlled biochemical activities, protein-protein interactions, protein-nucleic acid interactions, and protein-small molecule interactions. This unit presents a protocol for constructing functional yeast protein microarrays for global kinase substrate identification. This approach enables the rapid determination of protein interaction networks in yeast on a proteome-wide level. The same methodology can be readily applied to higher eukaryotic systems with careful consideration of overexpression strategy.
authors.affiliation: Department of Genetics, Stanford University, Stanford, California, USA.
authors.forename: Hogune
authors.initials: H
authors.lastname: Im
authors.forename: Michael
authors.initials: M
authors.lastname: Snyder
mesh_terms: D000818:Animals; D005656:Fungal Proteins / Q000032:analysis / Q000378:metabolism; D006801:Humans; D010766:Phosphorylation; D010770:Phosphotransferases / Q000032:analysis* / Q000378:metabolism; D040081:Protein Array Analysis / Q000379:methods*; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D020543:Proteome; D011994:Recombinant Proteins; D010770:Phosphotransferases
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