title: Identification of SUMO E3 ligase-specific substrates using the HuProt human proteome microarray.
abstract: The functional protein microarray is a powerful and versatile systems biology and proteomics tool that allows the rapid activity profiling of thousands of proteins in parallel. We have recently developed a human proteome array, the HuProt array, which includes ~80 % of all the full-length proteins of the human proteome. In one recent application of the HuProt array, we identified numerous SUMO E3 ligase-dependent SUMOylation substrates. For many SUMO E3 ligases, only a small number of substrates have been identified and the target specificities of these ligases therefore remain poorly defined. In this protocol, we outline a method we developed using the HuProt array to screen the human proteome to identify novel SUMO E3 ligase substrates recognized by specific E3 ligases.
authors.affiliation: Biochemistry, Cellular and Molecular Biology Graduate Program, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Cox
authors.forename: Ijeoma
authors.initials: I
authors.lastname: Uzoma
authors.forename: Catherine
authors.initials: C
authors.lastname: Guzzo
authors.forename: Jun Seop
authors.initials: JS
authors.lastname: Jeong
authors.forename: Michael
authors.initials: M
authors.lastname: Matunis
authors.forename: Seth
authors.initials: S
authors.lastname: Blackshaw
authors.forename: Heng
authors.initials: H
authors.lastname: Zhu
mesh_terms: D006801:Humans; D040081:Protein Array Analysis / Q000379:methods*; D011499:Protein Processing, Post-Translational; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D025841:Small Ubiquitin-Related Modifier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity; D058207:Sumoylation; D044767:Ubiquitin-Protein Ligases / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D020543:Proteome; D025841:Small Ubiquitin-Related Modifier Proteins; D044767:Ubiquitin-Protein Ligases
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