Title:
PROTEIN SUITABLE FOR PROTEIN ORIENTATION CONTROL/IMMOBILIZATION AND IMMOBILIZATION SUPPORT FOR THE PROTEIN
Document Type and Number:
WIPO Patent Application WO/2008/044692
Kind Code:
A1
Abstract:
A novel protein modified so as to attain specific highly efficient linkage of protein via its carboxyl terminal to an immobilization support, having the amino acid sequence of the general formula: R1-R2-R3-R4-R5 wherein the sequence starts with an amino terminal and ends with a carboxyl terminal, the sequence of R1 segment is a sequence of immobilization object protein, characterized by not containing lysine and cysteine residues; the sequence of R2 segment can be absent and when being present is a spacer sequence composed of amino acid residues other than lysine and cysteine residues; the sequence of R3 segment is a sequence composed of two amino acid residues expressed as cysteine-X (X is an amino acid residue other than lysine and cysteine residues); the sequence of R4 segment can be absent and when being present is a sequence not containing lysine and cysteine residues, characterized by containing an acidic amino acid residue capable of causing the isoelectric point of the protein of the amino acid sequence of the general formula R1-R2-R3-R4-R5 as a whole to fall in the acid side; and the sequence of R5 segment is an affinity tag sequence for protein purification. In the protein, the R1-R2 segments are used for immobilization on an immobilization support.
Inventors:
IWAKURA MASAHIRO (JP)
HIROTA KIYONORI (JP)
SOTA HIROYUKI (JP)
SARARA GOU (JP)
ARUGA YUKIKO (JP)
YAMANE CHIORI (JP)
HIROTA KIYONORI (JP)
SOTA HIROYUKI (JP)
SARARA GOU (JP)
ARUGA YUKIKO (JP)
YAMANE CHIORI (JP)
Application Number:
PCT/JP2007/069722
Publication Date:
April 17, 2008
Filing Date:
October 10, 2007
Export Citation:
Assignee:
NAT INST OF ADVANCED IND SCIEN (JP)
IWAKURA MASAHIRO (JP)
HIROTA KIYONORI (JP)
SOTA HIROYUKI (JP)
SARARA GOU (JP)
ARUGA YUKIKO (JP)
YAMANE CHIORI (JP)
IWAKURA MASAHIRO (JP)
HIROTA KIYONORI (JP)
SOTA HIROYUKI (JP)
SARARA GOU (JP)
ARUGA YUKIKO (JP)
YAMANE CHIORI (JP)
International Classes:
C07K14/00; C12N15/09; C07K17/00
Domestic Patent References:
| WO2005035585A1 | 2005-04-21 | |||
| WO2001000797A1 | 2001-01-04 |
Foreign References:
| JP2005058059A | 2005-03-10 |
Other References:
TAKENAWA T. ET AL.: "Cyanocysteine-mediated molecular dissection of dihydrofolate reductase: occurrence of intra- and inter-molecular reactions forming a peptide bond", J. BIOCHEM. (TOKYO), vol. 123, no. 6, 1998, pages 1137 - 1144, XP009018697
IWAKURA M. ET AL.: "Evolutional design of a hyperactive cysteine- and methionine-free mutant of Escherichia coli dihydrofolate reductase", J. BIOL. CHEM., vol. 281, no. 19, 2006, pages 13234 - 13246, XP003022197
IWAKURA M. ET AL.: "Evolutional design of a hyperactive cysteine- and methionine-free mutant of Escherichia coli dihydrofolate reductase", J. BIOL. CHEM., vol. 281, no. 19, 2006, pages 13234 - 13246, XP003022197
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