1. Characterization and structure of glyceraldehyde-3-phosphate dehydrogenase type 1 from Escherichia coli
- Author
-
I. K. Bostrom, Y. C. Yao, A. Q. Chen, S. H. Gu, Li Zhang, Meiruo Liu, Chaoneng Ji, Yuandong Wang, and Jie Li
- Subjects
Models, Molecular ,Protein Conformation, alpha-Helical ,Genetic Vectors ,Biophysics ,Gene Expression ,Shigella sonnei ,Dehydrogenase ,Crystallography, X-Ray ,medicine.disease_cause ,Glyceraldehyde 3-Phosphate ,Biochemistry ,Research Communications ,03 medical and health sciences ,Structural Biology ,Catalytic Domain ,Escherichia coli ,Genetics ,medicine ,Humans ,Protein Interaction Domains and Motifs ,Glycolysis ,Amino Acid Sequence ,Cloning, Molecular ,Glyceraldehyde 3-phosphate dehydrogenase ,030304 developmental biology ,Thermostability ,chemistry.chemical_classification ,0303 health sciences ,biology ,Escherichia coli Proteins ,030302 biochemistry & molecular biology ,Glyceraldehyde-3-Phosphate Dehydrogenases ,NAD ,Condensed Matter Physics ,Recombinant Proteins ,Protein Subunits ,Enzyme ,chemistry ,biology.protein ,Protein Conformation, beta-Strand ,NAD+ kinase ,Protein Binding - Abstract
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key enzyme in the glycolytic pathway that catalyzes the conversion of D-glyceraldehyde 3-phosphate to 1,3-diphosphoglycerate. Here, the full-length GAPDH type 1 from Escherichia coli (EcGAPDH1) was cloned and overexpressed, and the protein was purified. Biochemical analyses found that the optimum reaction temperature and pH of EcGAPDH1 were 55°C and 10.0, respectively. The protein has a certain amount of thermostability. Crystals of EcGAPDH1 were obtained using the sitting-drop vapor-diffusion technique and X-ray diffraction data were collected to 1.88 Å resolution. Characterization of the crystals showed that they belonged to space group P41212, with unit-cell parameters a = b = 89.651, c = 341.007 Å, α = β = γ = 90°. The structure of EcGAPDH1 contains four subunits, each of which includes an N-terminal NAD+-binding domain and a C-terminal catalytic domain. Analysis of the NAD+-bound form showed some differences between the structures of EcGAPDH1 and human GAPDH. As EcGAPDH1 shares 100% identity with GAPDH from Shigella sonnei, its structure may help in finding a drug for the treatment of shigellosis.
- Published
- 2020