5KJZ

Co-crystal structure of PKA RI alpha CNB-B mutant (G316R/A336T) with cGMP

5KJZ の概要

• エントリーDOI: 10.2210/pdb5kjz/pdb
• 関連するPDBエントリー: 5KJX 5KJY
• 分子名称: cAMP-dependent protein kinase type I-alpha regulatory subunit, CYCLIC GUANOSINE MONOPHOSPHATE, GLYCEROL, ... (4 entities in total)
• 機能のキーワード: cyclic nucleotide, camp-dependent protein kinase, nucleotide selectivity, cyclic nucleotide binding domain, signaling protein
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 17616.06

構造登録者

Lorenz, R.,Moon, E.,Kim, J.J.,Huang, G.Y.,Kim, C.,Herberg, F.W. (登録日: 2016-06-20, 公開日: 2017-06-28, 最終更新日: 2024-03-06)

主引用文献

Lorenz, R.,Moon, E.W.,Kim, J.J.,Schmidt, S.H.,Sankaran, B.,Pavlidis, I.V.,Kim, C.,Herberg, F.W.
Mutations of PKA cyclic nucleotide-binding domains reveal novel aspects of cyclic nucleotide selectivity.
Biochem. J., 474:2389-2403, 2017

PubMed Abstract: Cyclic AMP and cyclic GMP are ubiquitous second messengers that regulate the activity of effector proteins in all forms of life. The main effector proteins, the 3',5'-cyclic adenosine monophosphate (cAMP)-dependent protein kinase (PKA) and the 3',5'-cyclic guanosine monophosphate (cGMP)-dependent protein kinase (PKG), are preferentially activated by cAMP and cGMP, respectively. However, the molecular basis of this cyclic nucleotide selectivity is still not fully understood. Analysis of isolated cyclic nucleotide-binding (CNB) domains of PKA regulatory subunit type Iα (RIα) reveals that the C-terminal CNB-B has a higher cAMP affinity and selectivity than the N-terminal CNB-A. Here, we show that introducing cGMP-specific residues using site-directed mutagenesis reduces the selectivity of CNB-B, while the combination of two mutations (G316R/A336T) results in a cGMP-selective binding domain. Furthermore, introducing the corresponding mutations (T192R/A212T) into the PKA RIα CNB-A turns this domain into a highly cGMP-selective domain, underlining the importance of these contacts for achieving cGMP specificity. Binding data with the generic purine nucleotide 3',5'-cyclic inosine monophosphate (cIMP) reveal that introduced arginine residues interact with the position 6 oxygen of the nucleobase. Co-crystal structures of an isolated CNB-B G316R/A336T double mutant with either cAMP or cGMP reveal that the introduced threonine and arginine residues maintain their conserved contacts as seen in PKG I CNB-B. These results improve our understanding of cyclic nucleotide binding and the molecular basis of cyclic nucleotide specificity.

PubMed: 28583991
DOI: 10.1042/BCJ20160969

実験手法

X-RAY DIFFRACTION (1.347 Å)