TY  - JOUR
A1  - Möbius, Kristin
A1  - Kazemi, Sina
A1  - Güntert, Peter
A1  - Jakob, Andreas
A1  - Heckel, Alexander
A1  - Becker-Baldus, Johanna
A1  - Glaubitz, Clemens
T1  - Global response of diacylglycerol kinase towards substrate binding observed by 2D and 3D MAS NMR
T2  - Scientific reports
N2  - Escherichia coli diacylglycerol kinase (DGK) is an integral membrane protein, which catalyses the ATP-dependent phosphorylation of diacylglycerol (DAG) to phosphatic acid (PA). It is a unique trimeric enzyme, which does not share sequence homology with typical kinases. It exhibits a notable complexity in structure and function despite of its small size. Here, chemical shift assignment of wild-type DGK within lipid bilayers was carried out based on 3D MAS NMR, utilizing manual and automatic analysis protocols. Upon nucleotide binding, extensive chemical shift perturbations could be observed. These data provide evidence for a symmetric DGK trimer with all of its three active sites concurrently occupied. Additionally, we could detect that the nucleotide substrate induces a substantial conformational change, most likely directing DGK into its catalytic active form. Furthermore, functionally relevant interprotomer interactions are identified by DNP-enhanced MAS NMR in combination with site-directed mutagenesis and functional assays.
KW  - Kinases
KW  - Molecular conformation
KW  - Solid-state NMR
Y1  - 2019
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/48480
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-484807
SN  - 2045-2322
N1  - Open Access: This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
VL  - 9
IS  - 1, Art. 3995
SP  - 1
EP  - 17
PB  - Macmillan Publishers Limited, part of Springer Nature
CY  - [London]
ER  -