Title:
METHOD FOR IDENTIFYING POLYUBIQUITINATED SUBSTRATE
Document Type and Number:
WIPO Patent Application WO/2015/072507
Kind Code:
A1
Abstract:
The present invention provides a method for efficiently identifying a polyubiquitinated substrate, which is generally difficult to identify. This method for identifying a polyubiquitinated substrate has (1) a step for causing a trypsin-resistant polyubiquitin chain binding protein and ubiquitin ligase to be expressed in a cell, (2) a step for separating a complex containing the trypsin-resistant polyubiquitin chain binding protein from the cell subsequent to step (1), (3) a step for trypsin-digesting the complex separated by step (2), and (4) a step for identifying a peptide including a ubiquitinated site from the digest obtained by step (3).
Inventors:
YOSHIDA YUKIKO (JP)
SAEKI YASUSHI (JP)
TSUCHIYA HIKARU (JP)
MURAKAMI ARISA (JP)
TANAKA KEIJI (JP)
SAEKI YASUSHI (JP)
TSUCHIYA HIKARU (JP)
MURAKAMI ARISA (JP)
TANAKA KEIJI (JP)
Application Number:
PCT/JP2014/080053
Publication Date:
May 21, 2015
Filing Date:
November 13, 2014
Export Citation:
Assignee:
TOKYO METROPOLITAN INST MEDICAL SCIENCE (JP)
International Classes:
C12Q1/527; C12Q1/37; C12N15/09
Other References:
KIYOTAKA OSHIKAWA ET AL.: "Comprehensive study of protein ubiquitylation sites by conjugation of engineered lysine-less ubiquitin", JOURNAL OF JAPANESE BIOCHEMICAL SOCIETY, vol. 84, no. 6, 2012, pages 479 - 487, XP008183673
LOPITZ-OTSOA, F. ET AL.: "Integrative analysis of the ubiquitin proteome isolated using Tandem Ubiquitin Binding Entities (TUBEs", J. PROTEOMICS, vol. 75, 2012, pages 2998 - 3014, XP028923452
HJERPE, R. ET AL.: "Efficient protection and isolation of ubiquitylated proteins using tandem ubiquitin-binding entities", EMBO REPORTS, vol. 10, no. 11, 2009, pages 1250 - 1258, XP055341878
SHI, Y. ET AL.: "A Data Set of Human Endogenous Protein Ubiquitination Sites", MOLECULAR & CELLULAR PROTEOMICS, vol. 10, no. 5, 2011, XP055341879
YUKIKO YOSHIDA ET AL.: "Polyubiquitin Ketsugo Tanpakushitsu o Riyo shita Polyubiquitin-ka Kishitsu no Koritsuteki Doteiho no Kakuritsu", ANNUAL MEETING OF THE MOLECULAR BIOLOGY SOCIETY OF JAPAN PROGRAM YOSHISHU, 20 November 2013 (2013-11-20), XP008183998
KOMANDER; RAPE, ANNUAL REVIEW OF BIOCHEMISTRY, vol. 81, 2012, pages 203 - 229
GRABBE ET AL., NATURE REVIEWS. MOLECULAR CELL BIOLOGY, vol. 12, 2011, pages 295 - 307
HJERPE ET AL., EMBO REPORTS, vol. 10, 2009, pages 1250 - 1258
KIM ET AL., MOLECULAR CELL, vol. 44, 2011, pages 325 - 340
FRESCAS; PAGANO, NATURE REVIEWS CANCER, vol. 8, 2008, pages 438 - 449
LOPITZ-OTSOA, F. ET AL.: "Integrative analysis of the ubiquitin proteome isolated using Tandem Ubiquitin Binding Entities (TUBEs", J. PROTEOMICS, vol. 75, 2012, pages 2998 - 3014, XP028923452
HJERPE, R. ET AL.: "Efficient protection and isolation of ubiquitylated proteins using tandem ubiquitin-binding entities", EMBO REPORTS, vol. 10, no. 11, 2009, pages 1250 - 1258, XP055341878
SHI, Y. ET AL.: "A Data Set of Human Endogenous Protein Ubiquitination Sites", MOLECULAR & CELLULAR PROTEOMICS, vol. 10, no. 5, 2011, XP055341879
YUKIKO YOSHIDA ET AL.: "Polyubiquitin Ketsugo Tanpakushitsu o Riyo shita Polyubiquitin-ka Kishitsu no Koritsuteki Doteiho no Kakuritsu", ANNUAL MEETING OF THE MOLECULAR BIOLOGY SOCIETY OF JAPAN PROGRAM YOSHISHU, 20 November 2013 (2013-11-20), XP008183998
KOMANDER; RAPE, ANNUAL REVIEW OF BIOCHEMISTRY, vol. 81, 2012, pages 203 - 229
GRABBE ET AL., NATURE REVIEWS. MOLECULAR CELL BIOLOGY, vol. 12, 2011, pages 295 - 307
HJERPE ET AL., EMBO REPORTS, vol. 10, 2009, pages 1250 - 1258
KIM ET AL., MOLECULAR CELL, vol. 44, 2011, pages 325 - 340
FRESCAS; PAGANO, NATURE REVIEWS CANCER, vol. 8, 2008, pages 438 - 449
Attorney, Agent or Firm:
TANAI Sumio et al. (JP)
Sumio Tanai (JP)
Sumio Tanai (JP)
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