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Characterization and structure of glyceraldehyde‐3‐phosphate dehydrogenase type 1 from Escherichia coli
Author(s) -
Zhang L.,
Liu M. R.,
Yao Y. C.,
Bostrom I. K.,
Wang Y. D.,
Chen A. Q.,
Li J. X.,
Gu S. H.,
Ji C. N.
Publication year - 2020
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x20010067
Subject(s) - glyceraldehyde 3 phosphate dehydrogenase , thermostability , dehydrogenase , escherichia coli , nad+ kinase , biochemistry , chemistry , enzyme , biology , crystallography , gene
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 ( Ec GAPDH1) was cloned and overexpressed, and the protein was purified. Biochemical analyses found that the optimum reaction temperature and pH of Ec GAPDH1 were 55°C and 10.0, respectively. The protein has a certain amount of thermostability. Crystals of Ec GAPDH1 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 P 4 1 2 1 2, with unit‐cell parameters a = b = 89.651, c = 341.007 Å, α = β = γ = 90°. The structure of Ec GAPDH1 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 Ec GAPDH1 and human GAPDH. As Ec GAPDH1 shares 100% identity with GAPDH from Shigella sonnei , its structure may help in finding a drug for the treatment of shigellosis.