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8hrq

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Crystal structure of glyceraldehyde-3-phosphate dehydrogenase from Corynebacterium glutamicum ATCC13032 (L36S/T37K) in complex with NAD

Structural highlights

8hrq is a 2 chain structure with sequence from Corynebacterium glutamicum ATCC 13032. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:	X-ray diffraction, Resolution 2.09Å
Ligands:	, , , ,
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

G3P_CORGL Catalyzes the oxidative phosphorylation of glyceraldehyde 3-phosphate (G3P) to 1,3-bisphosphoglycerate (BPG) using the cofactor NAD. The first reaction step involves the formation of a hemiacetal intermediate between G3P and a cysteine residue, and this hemiacetal intermediate is then oxidized to a thioester, with concomitant reduction of NAD to NADH. The reduced NADH is then exchanged with the second NAD, and the thioester is attacked by a nucleophilic inorganic phosphate to produce BPG.[UniProtKB:P9WN83]

Publication Abstract from PubMed

Since the discovery of l-glutamate-producing Corynebacterium glutamicum, it has evolved to be an industrial workhorse. For biobased chemical production, suppling sufficient amounts of the NADPH cofactor is crucial. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a glycolytic enzyme that converts glyceraldehyde-3-phosphate (G3P) to 1,3-bisphosphoglycerate and produces NADH, is a major prospective solution for the cofactor imbalance issue. In this study, we determined the crystal structure of GAPDH from C. glutamicum ATCC13032 (CgGAPDH). Based on the structural information, we generated six CgGAPDH variants, CgGAPDH(L36S), CgGAPDH(L36S/T37K), CgGAPDH(L36S/T37K/P192S), CgGAPDH(L36S/T37K/F100V/P192S), CgGAPDH(L36S/T37K/F100L/P192S), and CgGAPDH(L36S/T37K/F100I/P192S), that can produce both NADH and NAPDH. The final CgGAPDH(L36S/T37K/F100V/P192S) variant showed a 212-fold increase in enzyme activity for NADP as well as 200% and 30% increased activity for the G3P substrate under NAD and NADP cofactor conditions, respectively. In addition, crystal structures of CgGAPDH variants in complex with NAD(P) permit the elucidation of differences between wild-type CgGAPDH and variants in relation to cofactor stabilization.

Structure-Guided Protein Engineering of Glyceraldehyde-3-phosphate Dehydrogenase from Corynebacterium glutamicum for Dual NAD/NADP Cofactor Specificity.,Son HF, Yu H, Hong J, Lee D, Kim IK, Kim KJ J Agric Food Chem. 2023 Nov 22;71(46):17852-17859. doi: 10.1021/acs.jafc.3c06176. , Epub 2023 Nov 7. PMID:37935620^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

↑ Son HF, Yu H, Hong J, Lee D, Kim IK, Kim KJ. Structure-Guided Protein Engineering of Glyceraldehyde-3-phosphate Dehydrogenase from Corynebacterium glutamicum for Dual NAD/NADP Cofactor Specificity. J Agric Food Chem. 2023 Nov 22;71(46):17852-17859. PMID:37935620 doi:10.1021/acs.jafc.3c06176