Identification of SUMO E3 ligase-specific substrates using the HuProt human proteome microarray

Eric Cox, Ijeoma Uzoma, Catherine Guzzo, Jun Seop Seop Jeong, Michael Matunis, Seth Blackshaw, Heng Zhu

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)455-463
Number of pages9
JournalMethods in Molecular Biology
Volume1295
DOIs
StatePublished - 2015

Keywords

  • E3 ligases
  • Microarray
  • Posttranslational modifications
  • Proteomics
  • SUMO

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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