SEQUENCING METHOD AND APPARATUS OF AMINO ACID OF PROTEIN

• Main Authors: TANIGUCHI KENICHI, YOSHIZAWA AKIYASU
• Format: Patent
• Language: English; Japanese
• Published: 24.08.2015
• Subjects: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; PHYSICS; TESTING

PROBLEM TO BE SOLVED: To accurately estimate the sequence of the terminal portion of a protein and also improve the reliability of the sequence estimation of other parts than the terminal parts even in the case of using protein DB based on the cDNA.SOLUTION: Based on the amino acid sequence on the protein DB based on the cDNA, multiple amino acid sequences from each end of which one amino acid residue is removed is created among the amino acid sequences of a predetermined length from the distal end of the terminal portion for each protein and a terminal sequence DB is created in association with each protein. Based on the result of the MSanalysis of the peptide mixture derived from the target protein, the MS/MS ion search using the terminal sequence DB is performed. Since, on the terminal sequence DB, the sequence which matches the amino acid sequence of the terminal portion of the measured protein from which a signal peptide is left out is recorded, the terminal peptide can be also identified and it is possible to identify the whole protein by matching the search results using the Swiss-Prot. 【課題】ｃＤＮＡに基づくタンパク質ＤＢを使用した場合でも、タンパク質の末端部分の配列を的確に推定するとともに末端部分以外の配列推定の信頼性も向上させる。【解決手段】ｃＤＮＡに基づくタンパク質ＤＢに収録されているアミノ酸配列に基づき、タンパク質毎に、末端部分の最末端から所定長さのアミノ酸配列の中で最末端からアミノ酸残基を１個ずつ除去した複数のアミノ酸配列を作成してタンパク質に対応付けた末端配列ＤＢを作成しておく。そして、目的タンパク質由来のペプチド混合物をＭＳ2分析した結果に基づき、末端配列ＤＢを使用したＭＳ／ＭＳイオンサーチを行う。末端配列ＤＢには、シグナルペプチドが脱落した実測のタンパク質の末端部分のアミノ酸配列と合致する配列が収録されているので、末端ペプチドも同定することができ、Swiss-Protを使用した検索結果と合わせることでタンパク質全体を同定することができる。【選択図】図３