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

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

Research output: Contribution to journalArticle

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 (Clifton, N.J.)
Volume1295
DOIs
StatePublished - 2015

Fingerprint

Ubiquitin-Protein Ligases
Proteome
Sumoylation
Protein Array Analysis
Systems Biology
Ligases
Proteomics
Proteins

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Identification of SUMO E3 ligase-specific substrates using the HuProt human proteome microarray. / Cox, Eric; Uzoma, Ijeoma; Guzzo, Catherine; Jeong, Jun Seop; Matunis, Michael; Blackshaw, Seth; Zhu, Heng.

In: Methods in molecular biology (Clifton, N.J.), Vol. 1295, 2015, p. 455-463.

Research output: Contribution to journalArticle

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