Abstract Enzymes sample multiple conformations during their catalytic cycles.Chemical shifts from Nuclear Magnetic Resonance (NMR) are hypersensitive to conformational changes and ensembles in solution.Phosphomannomutase/phosphoglucomutase (PMM/PGM) is a ubiquitous four-domain enzyme that catalyzes phosphoryl transfer across phosphohexose substrates.
We compared states the enzyme visits during its catalytic cycle.Collective responses of Pseudomonas PMM/PGM to phosphosugar substrates and inhibitor were assessed using NMR-detected titrations.Affinities were estimated from binding isotherms obtained by principal component analysis (PCA).
Relationships among phosphosugar-enzyme associations emerge from PCA comparisons of the titrations.COordiNated Chemical Shifts bEhavior (CONCISE) Reducing High Energy Demand Associated with Air-Conditioning Needs in Saudi Arabia analysis provides novel discrimination of three ligand-bound states of PMM/PGM harboring a mutation that suppresses activity.Enzyme phosphorylation and phosphosugar binding appear to drive the open dephosphorylated enzyme to the free phosphorylated state, and on toward ligand-closed states.
Domain 4 appears central to collective responses to substrate and inhibitor binding.Hydrogen exchange reveals that binding of a substrate analogue enhances folding stability of the domains to a uniform level, establishing a globally unified structure.CONCISE and PCA of NMR spectra have discovered novel states of a well-studied enzyme and appear ready to discriminate other enzyme and Essential Preventive Automobile Maintenance during a Pandemic ligand binding states.