Posttranslational modification assays on functional protein microarrays

Johnathan Neiswinger, Ijeoma Uzoma, Eric Cox, Heesool Rho, Jun Seop Jeong, Heng Zhu

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)892-899
Number of pages8
JournalCold Spring Harbor Protocols
Volume2016
Issue number10
DOIs
StatePublished - Oct 1 2016

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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