PMID:24032678

Citation	Arutyunov, D, Schmalhausen, E, Orlov, V, Rahuel-Clermont, S, Nagradova, N, Branlant, G and Muronetz, V (2013) An unusual effect of NADP(+) on the thermostability of the nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase from Streptococcus mutans. Biochem. Cell Biol. 91:295-302
Abstract	Adiabatic differential scanning calorimetry was used to investigate the effect of NADP(+) on the irreversible thermal denaturation of the nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPN) from Streptococcus mutans. The GAPN-NADP(+) binary complex showed a strongly decreased thermal stability, with a difference of about 20 °C between the temperatures of the thermal transition peak maxima of the complex and the free protein. This finding was similar to the previously described thermal destabilization of GAPN upon binding of inorganic phosphate to the substrate binding site and can be interpreted as the shift of the equilibrium between 2 conformers of tetrameric GAPN upon addition of the coenzyme. Single amino acid substitution, known to abolish the NADP(+) binding, cancelled the calorimetric effect of the coenzyme. GAPN thermal inactivation was considerably decelerated in the presence of NADP(+) showing that the apparent change in stability of the active centre can be the opposite to that of the whole protein molecule. NADP(+) could also reactivate the inactive GAPN* species, obtained by the heating of the apoenzyme below the thermal denaturation transition temperature. These effects may reflect a mechanism that provides GAPN the sufficient flexibility for the earlier observed profound active site reorganizations required during the catalytic cycle. The elevated thermal stability of the apoenzyme may, in turn, be important for maintaining a constant level of active GAPN - an enzyme that is known to be crucial for the effective supply of the reducing equivalents in S. mutans and its ability to grow under aerobic conditions.
Links	PubMed Online version:10.1139/bcb-2012-0104
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
STRMU:GAPN	enables	GO:0047100: glyceraldehyde-3-phosphate dehydrogenase (NADP+) (phosphorylating) activity	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
STRMU:GAPN		GO:0047100: glyceraldehyde-3-phosphate dehydrogenase (NADP+) (phosphorylating) activity	ECO:0000314:		F		Fig. 3 shows NADP+-induced restoration catalytic activity after thermal inactivation.	complete CACAO 8698

References

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