5JBN

Crystal Structure of Apo Phosphopantetheine Adenylyltransferase (PPAT/CoaD) from E. coli

5JBN の概要

• エントリーDOI: 10.2210/pdb5jbn/pdb
• 分子名称: Phosphopantetheine adenylyltransferase, SULFATE ION, DIMETHYL SULFOXIDE, ... (4 entities in total)
• 機能のキーワード: coad, apo, ppat, transferase
• 由来する生物種: Escherichia coli (strain K12)
• 細胞内の位置: Cytoplasm: P0A6I6
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 39146.76

構造登録者

Mamo, M.,Proudfoot, A.,Bussiere, D. (登録日: 2016-04-13, 公開日: 2016-05-25, 最終更新日: 2024-03-06)

主引用文献

Proudfoot, A.,Frank, A.O.,Ruggiu, F.,Mamo, M.,Lingel, A.
Facilitating unambiguous NMR assignments and enabling higher probe density through selective labeling of all methyl containing amino acids.
J.Biomol.Nmr, 65:15-27, 2016

PubMed Abstract: The deuteration of proteins and selective labeling of side chain methyl groups has greatly enhanced the molecular weight range of proteins and protein complexes which can be studied using solution NMR spectroscopy. Protocols for the selective labeling of all six methyl group containing amino acids individually are available, however to date, only a maximum of five amino acids have been labeled simultaneously. Here, we describe a new methodology for the simultaneous, selective labeling of all six methyl containing amino acids using the 115 kDa homohexameric enzyme CoaD from E. coli as a model system. The utility of the labeling protocol is demonstrated by efficiently and unambiguously assigning all methyl groups in the enzymatic active site using a single 4D (13)C-resolved HMQC-NOESY-HMQC experiment, in conjunction with a crystal structure. Furthermore, the six fold labeled protein was employed to characterize the interaction between the substrate analogue (R)-pantetheine and CoaD by chemical shift perturbations, demonstrating the benefit of the increased probe density.

PubMed: 27130242
DOI: 10.1007/s10858-016-0032-2

実験手法

X-RAY DIFFRACTION (1.45 Å)