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 language | English |
|---|---|
| Pages (from-to) | 3004-3007 |
| Number of pages | 4 |
| Journal | Proteomics |
| Volume | 8 |
| Issue number | 15 |
| DOIs | |
| Publication status | Published - Aug 1 2008 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - A parallel affinity purification method for selective isolation of polyubiquitinated proteins
AU - Ota, Kazuhisa
AU - Kito, Keiji
AU - Iemura, Shun Ichiro
AU - Natsume, Tohru
AU - Ito, Takashi
PY - 2008/8/1
Y1 - 2008/8/1
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=49749126346&partnerID=8YFLogxK
U2 - 10.1002/pmic.200800271
DO - 10.1002/pmic.200800271
M3 - Article
VL - 8
SP - 3004
EP - 3007
JO - Proteomics
JF - Proteomics
SN - 1615-9853
IS - 15
ER -