A parallel affinity purification method for selective isolation of polyubiquitinated proteins

Kazuhisa Ota, Keiji Kito, Shun Ichiro Iemura, Tohru Natsume, Takashi Ito

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

We developed a parallel affinity purification (PAP) procedure, in which ubiquitinated proteins are purified from the cells that coexpress two affinity-tagged ubiquitins by sequential use of affinity chromatography specific to each tag. In contrast with previous procedures using a single affinity-tagged ubiquitin, the PAP eliminates highly abundant ubiquitin monomers and monoubiquitinated proteins to selectively enrich proteins bearing both affinity-tags, or poly- and multiubiquitinated proteins. Accordingly, it would serve as a powerful method to facilitate mass-spectrometric identification of ubiquitinated proteins.

Original languageEnglish
Pages (from-to)3004-3007
Number of pages4
JournalProteomics
Volume8
Issue number15
DOIs
Publication statusPublished - Aug 1 2008
Externally publishedYes

Fingerprint

Ubiquitinated Proteins
Purification
Ubiquitin
Bearings (structural)
Ubiquitins
Affinity chromatography
Proteins
Affinity Chromatography
Monomers

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

A parallel affinity purification method for selective isolation of polyubiquitinated proteins. / Ota, Kazuhisa; Kito, Keiji; Iemura, Shun Ichiro; Natsume, Tohru; Ito, Takashi.

In: Proteomics, Vol. 8, No. 15, 01.08.2008, p. 3004-3007.

Research output: Contribution to journalArticle

Ota, Kazuhisa ; Kito, Keiji ; Iemura, Shun Ichiro ; Natsume, Tohru ; Ito, Takashi. / A parallel affinity purification method for selective isolation of polyubiquitinated proteins. In: Proteomics. 2008 ; Vol. 8, No. 15. pp. 3004-3007.
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