title: Posttranslational Modification Assays on Functional Protein Microarrays.
abstract: Protein microarray technology provides a straightforward yet powerful strategy for identifying substrates of posttranslational modifications (PTMs) and studying the specificity of the enzymes that catalyze these reactions. Protein microarray assays can be designed for individual enzymes or a mixture to establish connections between enzymes and substrates. Assays for four well-known PTMs-phosphorylation, acetylation, ubiquitylation, and SUMOylation-have been developed and are described here for use on functional protein microarrays. Phosphorylation and acetylation require a single enzyme and are easily adapted for use on an array. The ubiquitylation and SUMOylation cascades are very similar, and the combination of the E1, E2, and E3 enzymes plus ubiquitin or SUMO protein and ATP is sufficient for in vitro modification of many substrates.
authors.affiliation: Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; The Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
authors.forename: Johnathan
authors.initials: J
authors.lastname: Neiswinger
authors.affiliation: Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; The Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
authors.forename: Ijeoma
authors.initials: I
authors.lastname: Uzoma
authors.affiliation: Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; The Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
authors.forename: Eric
authors.initials: E
authors.lastname: Cox
authors.affiliation: Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; The Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
authors.forename: HeeSool
authors.initials: H
authors.lastname: Rho
authors.affiliation: Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; The Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
authors.forename: Jun Seop
authors.initials: JS
authors.lastname: Jeong
authors.affiliation: Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; The Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287.
authors.forename: Heng
authors.initials: H
authors.lastname: Zhu
mesh_terms: D000107:Acetylation; D046228:Microarray Analysis / Q000379:methods*; D010766:Phosphorylation; D040081:Protein Array Analysis / Q000379:methods*; D011499:Protein Processing, Post-Translational; D058207:Sumoylation; D054875:Ubiquitination