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MUTANT PYRROLYSYL-TRNA SYNTHETASE, AND METHOD FOR PRODUCTION OF PROTEIN HAVING NON-NATURAL AMINO ACID INTEGRATED THEREIN BY USING THE SAME 実績あり

外国特許コード F130007279
整理番号 23000
掲載日 2013年4月8日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2008JP067029
国際公開番号 WO 2009038195
国際出願日 平成20年9月19日(2008.9.19)
国際公開日 平成21年3月26日(2009.3.26)
優先権データ
  • 特願2007-243574 (2007.9.20) JP
発明の名称 (英語) MUTANT PYRROLYSYL-TRNA SYNTHETASE, AND METHOD FOR PRODUCTION OF PROTEIN HAVING NON-NATURAL AMINO ACID INTEGRATED THEREIN BY USING THE SAME 実績あり
発明の概要(英語) Disclosed is a method for introducing a lysine derivative (particularly an N&egr;-benzyloxycarbonyl-lysine (Z-Lys) derivative) having a useful functional group such as a heavy atom, selenium, a reactive functional group, a fluorescent group or a crosslinker, which is suitable as a non-natural amino acid, into a desired protein in a site-specific manner. Also disclosed is a mutant pyrrolysyl-tRNA shnthetase which has the substitution of at least one amino acid residue selected from a tyrosine residue at position-306, a leucine residue at position-309 and a cysteine residue at position-348 each constituting a pyrrolidin-binding site in the amino acid sequence for pyrrolysyl-tRNA shnthetase depicted in SEQ ID NO:2, wherein the substitution of the amino acid residue is the substitution of a tyrosine residue at position-306 by a glycine or alanine residue, the substitution of a leucine residue at position-309 by a glycine or alanine residue, or the substitution of a cysteine residue at position-348 by a valine, serine or alanine residue.
従来技術、競合技術の概要(英語) BACKGROUND ART
At a desired position in the protein amino acid residues, of protein synthesis associated with normal (non-natural amino acid) other than the type 20 is substituted with an amino acid, non-natural amino acid is incorporated in the protein (allo-protein), the structure of the proteins, for functional analysis of proteins can be an effective means. Aminoacyl - tRNA synthetase various from a species of organism using the pair (aaRS) /tRNA, one or more 30 allo already synthesized protein. Many useful and most and has long been applied to the non-natural amino acid is introduced into the system, - tRNA synthetase (TyrRS) variants which tRNATyr and pair. This method is, eubacteria, archaebacteria and eukaryotic aaRS and is one of the groups 2, aminoacylated tRNA in each group but is, of the other groups cannot be aminoacylated tRNA (orthogonal) orthogonality of the key and the relationship to each other. For example, archaeal , of (Methanocaldococcus jannaschii) TyrRS/tRNATyr pair E. coli system, on the other hand E. coli TyrRS Bacillus, stearothermophilus tRNATyr mammalian cells in a system of the pair of orthogonal pair for, in these lines can be used in dilatation of the genetic code (for example, see patent document 1 and non-patent document 1).
On the other hand, , (Methanosarcina mazei) - tRNA synthetase (PylRS) derived from pyrrolidyl and tRNAPyl is, aaRS/tRNA pair in E. coli cells function as has the orthogonal properties (for example, see non-patent document 2). Further, the pair, in eukaryotic cells can be used in the genetic code expansion is reported (for example, see Patent Document 2). Pyrrolidine sterically bulky side chain (Pyrrolysine) having a lysine derivative. Wild-type PylRS is, in E. coli Nε -Boc-L-lysine tRNAPyl may be bonded to a (see Patent Document 2). In addition, and wild-type PylRS, ATP analog, pyrrolidine, or of a complex between X ray crystal structure have been reported (non-patent document 3, see 4 and 9).
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • RIKEN
  • 発明者(英語)
  • YOKOYAMA, Shigeyuki
  • SAKAMOTO, Kensaku
  • YANAGISAWA, Tatsuo
  • KOBAYASHI, Takatsugu
国際特許分類(IPC)
指定国 National States: AE AG AL AM AO AT AU AZ BA BB BG BH BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IS JP KE KG KM KN KP KR KZ LA LC LK LR LS LT LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PG PH PL PT RO RS RU SC SD SE SG SK SL SM ST SV SY TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ MD RU TJ TM
EPO: AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MT NL NO PL PT RO SE SI SK TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

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