7UD6

Designed Enzyme SH3-588 (Catechol O-methyltransferase catalytic domain and Src homology 3 binding domain fusion)

7UD6 の概要

• エントリーDOI: 10.2210/pdb7ud6/pdb
• 分子名称: Tyrosine-protein kinase Fyn,Catechol O-methyltransferase, S-ADENOSYL-L-HOMOCYSTEINE, POTASSIUM ION, ... (4 entities in total)
• 機能のキーワード: designed enzyme, comt, sh3, fusion protein, transferase
• 由来する生物種: Homo sapiens (human); 詳細
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 33057.52

構造登録者

Ongpipattanakul, C.,Nair, S.K. (登録日: 2022-03-18, 公開日: 2023-01-11, 最終更新日: 2023-10-25)

主引用文献

Park, R.,Ongpipattanakul, C.,Nair, S.K.,Bowers, A.A.,Kuhlman, B.
Designer installation of a substrate recruitment domain to tailor enzyme specificity.
Nat.Chem.Biol., 19:460-467, 2023

PubMed Abstract: Promiscuous enzymes that modify peptides and proteins are powerful tools for labeling biomolecules; however, directing these modifications to desired substrates can be challenging. Here, we use computational interface design to install a substrate recognition domain adjacent to the active site of a promiscuous enzyme, catechol O-methyltransferase. This design approach effectively decouples substrate recognition from the site of catalysis and promotes modification of peptides recognized by the recruitment domain. We determined the crystal structure of this novel multidomain enzyme, SH3-588, which shows that it closely matches our design. SH3-588 methylates directed peptides with catalytic efficiencies exceeding the wild-type enzyme by over 1,000-fold, whereas peptides lacking the directing recognition sequence do not display enhanced efficiencies. In competition experiments, the designer enzyme preferentially modifies directed substrates over undirected substrates, suggesting that we can use designed recruitment domains to direct post-translational modifications to specific sequence motifs on target proteins in complex multisubstrate environments.

PubMed: 36509904
DOI: 10.1038/s41589-022-01206-0

実験手法

X-RAY DIFFRACTION (2.59 Å)