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Kinetic and structural insights into enzymatic mechanism of succinic semialdehyde dehydrogenase from Cyanothece sp. ATCC51142
- Source :
- PLoS ONE, PLoS ONE, Vol 15, Iss 9, p e0239372 (2020)
- Publication Year :
- 2020
-
Abstract
- As a ubiquitous enzyme, succinic semialdehyde dehydrogenase contributes significantly in many pathways including the tricarboxylic acid cycle and other metabolic processes such as detoxifying the accumulated succinic semialdehyde and surviving in nutrient-limiting conditions. Here the cce4228 gene encoding succinic semialdehyde dehydrogenase from Cyanothece sp. ATCC51142 was cloned and the homogenous recombinant cce4228 protein was obtained by Ni-NTA affinity chromatography. Biochemical characterization revealed that cce4228 protein displayed optimal activity at presence of metal ions in basic condition. Although the binding affinity of cce4228 protein with NAD+ was about 50-fold lower than that of cce4228 with NADP+, the catalytic efficiency of cce4228 protein towards succinic semialdehyde with saturated concentration of NADP+ is same as that with saturated concentration of NAD+ as its cofactors. Meanwhile, the catalytic activity of cce4228 was competitively inhibited by succinic semialdehyde substrate. Kinetic and structural analysis demonstrated that the conserved Cys262 and Glu228 residues were crucial for the catalytic activity of cce4228 protein and the Ser157 and Lys154 residues were determinants of cofactor preference.
- Subjects :
- Models, Molecular
Protein Conformation
Biochemistry
Succinic semialdehyde
Substrate Specificity
chemistry.chemical_compound
0302 clinical medicine
Recombinant Protein Purification
Enzyme Chemistry
chemistry.chemical_classification
Synechococcus
0303 health sciences
Multidisciplinary
Crystallography
biology
Physics
Biochemical Cofactors
Condensed Matter Physics
Recombinant Proteins
Enzymes
Physical Sciences
Crystal Structure
Medicine
Succinate-Semialdehyde Dehydrogenase
Oxidoreductases
Research Article
food.ingredient
Protein Purification
Cyanothece
Science
Cyanobacteria
Research and Analysis Methods
Cofactor
03 medical and health sciences
food
Affinity chromatography
Solid State Physics
Amino Acid Sequence
Dehydrogenases
030304 developmental biology
Bacteria
Organisms
Substrate (chemistry)
Biology and Life Sciences
Proteins
NAD
Anabaena
Citric acid cycle
Kinetics
Enzyme
chemistry
Mutation
biology.protein
Enzymology
NAD+ kinase
030217 neurology & neurosurgery
NADP
Purification Techniques
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 15
- Issue :
- 9
- Database :
- OpenAIRE
- Journal :
- PloS one
- Accession number :
- edsair.doi.dedup.....192014ab050d11f75bbb4dc22d848ae5