Summary of project PR000941
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000941. The data can be accessed directly via it's Project DOI: 10.21228/M8296W This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Project ID: | PR000941 |
| Project DOI: | doi: 10.21228/M8296W |
| Project Title: | A redox-active switch in Fructosamine-3-kinases expands the regulatory repertoire of the protein kinase super-family |
| Project Type: | Structural Biology, enzyme characterization |
| Project Summary: | Aberrant regulation of metabolic kinases by altered redox homeostasis is a major contributing factor in aging and disease such as diabetes. However, the biochemical mechanisms by which metabolic kinases are regulated under oxidative stress is poorly understood. In this study, we demonstrate that the catalytic activity of a conserved family of Fructosamine-3-kinases (FN3Ks), which are evolutionarily related to eukaryotic protein kinases (ePKs), are regulated by redox-active cysteines in the kinase domain. By solving the crystal structure of FN3K homolog from Arabidopsis thaliana (AtFN3K), we demonstrate that it forms an unexpected strand-exchange dimer in which the ATP binding P-loop and adjoining beta strands are swapped between two chains in the dimer. This dimeric configuration is characterized by strained inter-chain disulfide bonds that stabilize the P-loop in an extended conformation. Mutational analysis and solution studies confirm that the strained disulfides function as redox “switches” to reversibly regulate FN3K activity and dimerization. Consistently, we find that human FN3K, which contains an equivalent P-loop Cys, is also redox-sensitive, whereas ancestral bacterial FN3K homologs, which lack a P-loop Cys, are not. Furthermore, CRISPR knockout of FN3K in human HepG2 cells results in significant upregulation of redox metabolites including glutathione. We propose that redox regulation evolved progressively in FN3Ks in response to changing cellular redox conditions. Our studies provide important new insights into the origin and evolution of redox regulation in the protein kinase superfamily and open new avenues for targeting human FN3K in diabetic complications. |
| Institute: | University of Georgia |
| Department: | Biochemistry and Molecular Biology |
| Laboratory: | Kannan |
| Last Name: | Kannan |
| First Name: | Natarajan |
| Address: | B122 Life Sciences Bldg. University of Georgia Athens, GA 30602 |
| Email: | nkannan@uga.edu |
| Phone: | 706-542-1334 |
| Funding Source: | MCB-1149106; R01GM114409 |
| Publications: | A redox-active switch in Fructosamine-3-kinases expands the regulatory repertoire of the protein kinase super-family |
| Contributors: | Safal Shrestha, Samiksha Katiyar, Carlos E. Sanz-Rodriquez, Nolan R. Kemppinen, Hyun W. Kim, Renuka Kadirvelraj, Charalampos Panagos, Neda Keyhaninejad, Maxwell Colonna, Pradeep Chopra, Dominic P. Byrne, Geert J. Boons, Esther V. Knaap, Patrick A. Eyers, Arthur S. Edison, Zachary A. Wood |
Summary of all studies in project PR000941
| Study ID | Study Title | Species | Institute | Analysis(* : Contains Untargted data) | Release Date | Version | Samples | Download(* : Contains raw data) |
|---|---|---|---|---|---|---|---|---|
| ST001375 | Fructosamine-3-kinase (FN3K) KO in HepG2 liver cancer cells | Homo sapiens | University of Georgia | NMR | 2020-07-07 | 1 | 20 | Uploaded data (424.9M)* |
