Title:
METHOD FOR SCREENING FOR PEPTIDE SUBSTRATE BY ADDRESSABLE POSITIONAL SCANNING TECHNIQUE
Document Type and Number:
WIPO Patent Application WO/2012/029965
Kind Code:
A1
Abstract:
The present invention provides a peptide substrate screening method which has high through-put performance, is inexpensive, and has high reaction efficiency and excellent general versatility. This peptide substrate screening method comprises the steps of: (a) reacting a substrate mixture with an enzyme protein, wherein the substrate mixture comprises multiple groups of candidate peptide substrates, candidate peptide substrates in each of the groups of the substrates have amino acid sequences that share some common amino acid residues, and a nucleic acid tag having a specified nucleotide sequence is bound to the terminal of each of the amino acid sequences (wherein the common amino acid residues and the nucleotide sequence of the nucleic acid tag are different among the groups of the substrates); (b) adding the substrate mixture which has been subjected to the reaction onto a base on which a nucleic acid capable of hybridizing with the nucleic acid tag has been immobilized at a predetermined position thereon, thereby immobilizing each of the groups of the substrates at the predetermined position on the base; and (c) detecting a group of substrates which has reacted with the enzyme protein.
Inventors:
KATAYAMA YOSHIKI (JP)
MORI TAKESHI (JP)
HATA AKITO (JP)
YAYAMA YOSHIHIRO (JP)
MORI TAKESHI (JP)
HATA AKITO (JP)
YAYAMA YOSHIHIRO (JP)
Application Number:
PCT/JP2011/070087
Publication Date:
March 08, 2012
Filing Date:
August 29, 2011
Export Citation:
Assignee:
UNIV KYUSHU NAT UNIV CORP (JP)
KATAYAMA YOSHIKI (JP)
MORI TAKESHI (JP)
HATA AKITO (JP)
YAYAMA YOSHIHIRO (JP)
KATAYAMA YOSHIKI (JP)
MORI TAKESHI (JP)
HATA AKITO (JP)
YAYAMA YOSHIHIRO (JP)
International Classes:
C12N15/09; C12Q1/48; C12Q1/68
Other References:
LJUNGDAHL, T. ET AL.: "Positional Scanning Peptide Libraries for Kinase Substrate Specificity Determinations: Straightforward and Reproducible Synthesis Using Pentafluorophenyl Esters.", J. COMB. CHEM., vol. 12, no. 5, 5 December 2010 (2010-12-05), pages 733 - 742
HUTTI, J.E. ET AL.: "A rapid method for determining protein kinase phosphorylation specificity.", NATURE METHODS, vol. 1, no. 1, October 2004 (2004-10-01), pages 27 - 29
MOK, J. ET AL.: "Deciphering protein kinase specificity through large-scale analysis of yeast phosphorylation site motifs.", SCI. SIGNAL., vol. 3, no. 109, 16 February 2010 (2010-02-16), pages 1/29 - 29/29
POUCHAIN, D. ET AL.: "A 10,000 member PNA- encoded peptide library for profiling tyrosine kinases.", ACS CHEM. BIOL., vol. 2, no. 12, 21 December 2007 (2007-12-21), pages 810 - 818
YOSHIHIRO YAYAMA ET AL.: "PNA Tag Peptide ni yoru Protein Kinase Kishitsu Array Screening-ho no Kaihatsu", THE JAPAN SOCIETY FOR ANALYTICAL CHEMISTRY NENKAI KOEN YOSHISHU, vol. 59, 9 January 2010 (2010-01-09), pages 317
HUTTI, J.E. ET AL.: "A rapid method for determining protein kinase phosphorylation specificity.", NATURE METHODS, vol. 1, no. 1, October 2004 (2004-10-01), pages 27 - 29
MOK, J. ET AL.: "Deciphering protein kinase specificity through large-scale analysis of yeast phosphorylation site motifs.", SCI. SIGNAL., vol. 3, no. 109, 16 February 2010 (2010-02-16), pages 1/29 - 29/29
POUCHAIN, D. ET AL.: "A 10,000 member PNA- encoded peptide library for profiling tyrosine kinases.", ACS CHEM. BIOL., vol. 2, no. 12, 21 December 2007 (2007-12-21), pages 810 - 818
YOSHIHIRO YAYAMA ET AL.: "PNA Tag Peptide ni yoru Protein Kinase Kishitsu Array Screening-ho no Kaihatsu", THE JAPAN SOCIETY FOR ANALYTICAL CHEMISTRY NENKAI KOEN YOSHISHU, vol. 59, 9 January 2010 (2010-01-09), pages 317
Attorney, Agent or Firm:
KOBAYASHI, Hiroshi et al. (JP)
Kobayashi 浩 (JP)
Kobayashi 浩 (JP)
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Claims: