Chemical biology using the protein-introduced unnatural lysine derivatives as probes

2015 Fiscal Year Final Research Report

• Project/Area Number: 24550203
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Chemistry related to living body
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Yanagisawa Tatsuo 国立研究開発法人理化学研究所, 横山構造生物学研究室, 研究員 (10450420)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Keywords: バイオテクノロジー / 非天然型アミノ酸 / タンパク質 / 翻訳後修飾 / ケミカルバイオロジー / tRNA / ピロリジン / ピロリジルtRNA合成酵素

Outline of Final Research Achievements

We developed a novel method to prepare histone proteins bearing multiple N(epsilon)-monomethyllysine residues at specified positions. By the combination of the Release factor 1 (RF1)-knockout (RFzero) Escherichia coli cells and the pair of tRNAPyl and pyrrolysyl-tRNA synthetase (PylRS), a tert-butyloxycarbonyl (Boc)-protected N(epsilon)-monomethyllysine analogue (BocKme1) is translationally incorporated into one or more positions specified with the UAG codon. The Boc groups on the protein are then removed to generate N(epsilon)-monomethyllysine residues. We installed N(epsilon) -monomethyllysine residues at positions 4, 9, 27, 36, and/or 79 of histone H3. The present method enables the installation of authentic N(epsilon)-monomethyllysines at multiple positions within a protein for large-scale production and will lead to better understanding of epigenetics research.

Free Research Field

化学生物学、構造生物化学