6K1R

Crystal structure of Ketopantoate reductase from Pseudomonas aeruginosa in complex with NAD+ and ketopantoate

Summary for 6K1R

• Entry DOI: 10.2210/pdb6k1r/pdb
• Descriptor: Probable 2-dehydropantoate 2-reductase, GLYCEROL, KETOPANTOATE, ... (5 entities in total)
• Functional Keywords: vitamin b5 biosynthesis pathway, redox cofactor, oxidoreductase
• Biological source: Pseudomonas aeruginosa PAO1
• Total number of polymer chains: 3
• Total formula weight: 107780.39

Authors

Khanppnavar, B.,Choudhury, A.,Datta, S. (deposition date: 2019-05-12, release date: 2020-05-13, Last modification date: 2023-11-22)

Primary citation

Choudhury, A.,Khanppnavar, B.,Datta, S.
Crystallographic and biophysical analyses of Pseudomonas aeruginosa ketopantoate reductase: Implications of ligand induced conformational changes in cofactor recognition.
Biochimie, 2021

PubMed Abstract: Ketopantoate reductases (KPRs) catalyse NADPH-dependent reduction of ketopantoate to pantoate, the rate-limiting step of pantothenate biosynthetic pathway. In our recent study, we showed KPRs are under dynamic evolutionary selection and highlighted the possible role of ordered substrate binding kinetics for cofactor selection. To further delineate this at molecular level, here, we perform X-ray crystallographic and biophysical analyses of KPR in presence of non-canonical cofactor NAD. In our structure, NAD was found to be highly dynamic in catalytic pocket of KPR, which could attain stable conformation only in presence of ketopantoate. Further, isothermal calorimetric (ITC) titrations showed that affinity of KPR for ketopantoate is higher in presence of NADP than in presence of NAD and lowest in absence of redox cofactors. In sum, our results clearly depict two modes of redox cofactor selections in KPRs, firstly by specific salt bridge interactions with unique phosphate moiety of NADP and secondly via ordered sequential heterotrophic cooperative binding of substrate ketopantoate.

PubMed: 34757166
DOI: 10.1016/j.biochi.2021.10.015

Experimental method

X-RAY DIFFRACTION (2.55 Å)