The interaction between Escherichia coli aspartokinase-homoserine dehydrogenase and 3-acetylpyridine-adenine dinucleotide phosphate (reduced), an analog of NADPH.

Müller, K; Garel, Jean‐Renaud

doi:10.1016/s0021-9258(17)43345-x

Cited by 3 publications

(1 citation statement)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It does not seem either to be limited by a conformational change occurring after the encounter step and having a large enough amplitude to be sensitive to the solvent viscosity. Therefore, it is likely that the reactivation of the dehydrogenase is related to minor structural rearrangements within a dimer and not to major relative displacements of the folded regions; these rearrangements could be similar to the subtle conformation changes of the protein that are apparently needed for a normal dehydrogenase activity in native AK-HDH (Müller & Garel, 1984c).…”

Section: The Slow Reactivationmentioning

confidence: 99%

Mechanism of renaturation of a large protein, aspartokinase-homoserine dehydrogenase

Vaucheret¹,

Signon²,

Bras³

et al. 1987

Biochemistry

View full text Add to dashboard Cite

The renaturation of aspartokinase-homoserine dehydrogenase and of some of its smaller fragments has been investigated after complete unfolding by 6 M guanidine hydrochloride. Fluorescence measurements show that a major folding reaction occurs rapidly (in less than a few seconds) after the protein has been transferred to native conditions and results in the shielding of the tryptophan residues from the aqueous solvent; this step also takes place in the fragments and probably corresponds to the independent folding of different segments along the polypeptide chain. The reappearance of the kinase activity, which is an index of the formation of "native" structure within a single chain, is much slower (a few minutes) and has the following properties: it is involved in a kinetic competition with the formation of aggregates; it has an activation energy of 22 +/- 5 kcal/mol; it is not related to a slow reaction in unfolding and thus probably not controlled by the cis-trans isomerization of X-Pro peptide bonds; its rate is inversely proportional to the solvent viscosity. It seems as if this reaction is limited by the mutual arrangement of the regions that have folded rapidly and independently. It is proposed that the mechanism where a fast folding of domains is followed by a slow pairing of folded domains could be generalized to other long chains composed of several domains; such a slow pairing of folded domains would correspond to a rate-limiting process specific to the renaturation of large proteins. The reappearance of the dehydrogenase activity measures the formation of a dimeric species. The dimerization can occur only after each chain has reached its "native" conformation.(ABSTRACT TRUNCATED AT 250 WORDS)

show abstract

Section: The Slow Reactivationmentioning

confidence: 99%

Mechanism of renaturation of a large protein, aspartokinase-homoserine dehydrogenase

Vaucheret¹,

Signon²,

Bras³

et al. 1987

Biochemistry

View full text Add to dashboard Cite

show abstract

Mutagenesis of Key Residues in the Binding Center of l‐Aspartate‐β‐Semialdehyde Dehydrogenase from Escherichia coli Enhances Utilization of the Cofactor NAD(H)

Wang

Duan

et al. 2015

ChemBioChem

View full text Add to dashboard Cite

L-Aspartate-β-semialdehyde dehydrogenase (ASADH) is a key enzyme in the aspartate pathway. In bacteria, ASADH is highly specific for the cofactor NADP(+) rather than NAD(+). Limited information on cofactor utilization is available, and neither the wild-type protein nor the available mutants could utilize NAD(+) efficiently. In this study, we identified several residues crucial for cofactor utilization by Escherichia coli ASADH (ecASADH) by mutating residues within the cofactor binding center. Among the investigated mutants, ecASADH-Q350N and ecASADH-Q350N/H171A, which exhibited markedly improved NAD(+) utilization, were further investigated by various biochemical approaches and molecular modeling. Relative to the wild type, the two mutants showed approximately 44-fold and 66-fold increases, respectively, in the constant kcat /Km of NAD(+). As desired, they could also utilize NADH efficiently to synthesize l-homoserine in cascade reactions in vitro.

show abstract

Characterization of the reduction of 3-acetylpyridine adenine dinucleotide phosphate by benzyl alcohol catalyzed by aldose reductase

Griffin

McNatt

1986

Archives of Biochemistry and Biophysics

View full text Add to dashboard Cite

The interaction between Escherichia coli aspartokinase-homoserine dehydrogenase and 3-acetylpyridine-adenine dinucleotide phosphate (reduced), an analog of NADPH.

Cited by 3 publications

References 14 publications

Mechanism of renaturation of a large protein, aspartokinase-homoserine dehydrogenase

Mechanism of renaturation of a large protein, aspartokinase-homoserine dehydrogenase

Mutagenesis of Key Residues in the Binding Center of l‐Aspartate‐β‐Semialdehyde Dehydrogenase from Escherichia coli Enhances Utilization of the Cofactor NAD(H)

Characterization of the reduction of 3-acetylpyridine adenine dinucleotide phosphate by benzyl alcohol catalyzed by aldose reductase

Contact Info

Product

Resources

About