Visualization of Nucleotidyl Transfer Reaction by Human DNA Polymerase η Using Time Resolved Protein Crystallography

• NAKAMURA Teruya

  Graduate School of Pharmaceutical Sciences, Kumamoto University

• YAMAGATA Yuriko

  Graduate School of Pharmaceutical Sciences, Kumamoto University

• YANG Wei

  Laboratory of Molecular Biology, NIDDK, NIH

Bibliographic Information

Other Title

• 時分割タンパク質結晶学によるDNAポリメラーゼηのヌクレオチド転移反応の可視化
• ジブンカツ タンパクシツ ケッショウガク ニ ヨル DNA ポリメラーゼe ノ ヌクレオチド テンイ ハンノウ ノ カシカ

Abstract

We visualized the course of nucleotidyl transfer reaction by human DNA polymerase η (Pol η) using time resolved protein crystallography. The reaction was initiated by exposing Pol η-DNA-dATP crystals to 1 mM Mg²⁺ at pH 7.0 and stopped at various time points by freezing. The substrates and two Mg²⁺ ions are aligned for reaction within 40 s. Transient electron densities indicate that deprotonation and an accompanying C2'-endo to C3'-endo conversion of the nucleophile 3'-OH are rate limiting. A third Mg²⁺ ion, which arrives with the new bond and stabilizes the intermediate state, may be an unappreciated feature of the two-metal-ion mechanism.

Journal

• Nihon Kessho Gakkaishi

  Nihon Kessho Gakkaishi 55 (1), 42-46, 2013

  The Crystallographic Society of Japan

Details

• CRID: 1390282679067175680
• NII Article ID: 10031156005
• NII Book ID: AN00188364
• DOI: 10.5940/jcrsj.55.42
• COI: 1:CAS:528:DC%2BC3sXnvVelt7o%3D
• ISSN: 18845576
• ISSN: 03694585
• NDL BIB ID: 024334927
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I024334927
• Text Lang: ja
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed