2VYV

Structure of E.Coli GAPDH Rat Sperm GAPDH heterotetramer

Summary for 2VYV

• Entry DOI: 10.2210/pdb2vyv/pdb
• Related: 1DC3 1DC4 1DC5 1DC6 1GAD 1GAE 1S7C 2VYN
• Descriptor: GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE, FORMIC ACID, NICOTINAMIDE-ADENINE-DINUCLEOTIDE, ... (6 entities in total)
• Functional Keywords: contraceptive, oxidoreductase, alpha chlorohydrin, glyceraldehyde-3-phosphate, nad, gapdh, rat sperm, cytoplasm, glycolysis
• Biological source: ESCHERICHIA COLI BL21(DE3); More
• Total number of polymer chains: 4
• Total formula weight: 146771.00

Authors

Frayne, J.,Taylor, A.,Hall, L.,Hadfield, A. (deposition date: 2008-07-29, release date: 2008-08-05, Last modification date: 2023-12-13)

Primary citation

Frayne, J.,Taylor, A.,Cameron, G.,Hadfield, A.T.
Structure of Insoluble Rat Sperm Glyceraldehyde-3-Phosphate Dehydrogenase (Gapdh) Via Heterotetramer Formation with Escherichia Coli Gapdh Reveals Target for Contraceptive Design.
J.Biol.Chem., 284:22703-, 2009

PubMed Abstract: Sperm glyceraldehyde-3-phosphate dehydrogenase has been shown to be a successful target for a non-hormonal contraceptive approach, but the agents tested to date have had unacceptable side effects. Obtaining the structure of the sperm-specific isoform to allow rational inhibitor design has therefore been a goal for a number of years but has proved intractable because of the insoluble nature of both native and recombinant protein. We have obtained soluble recombinant sperm glyceraldehyde-3-phosphate dehydrogenase as a heterotetramer with the Escherichia coli glyceraldehyde-3-phosphate dehydrogenase in a ratio of 1:3 and have solved the structure of the heterotetramer which we believe represents a novel strategy for structure determination of an insoluble protein. A structure was also obtained where glyceraldehyde 3-phosphate binds in the P(s) pocket in the active site of the sperm enzyme subunit in the presence of NAD. Modeling and comparison of the structures of human somatic and sperm-specific glyceraldehyde-3-phosphate dehydrogenase revealed few differences at the active site and hence rebut the long presumed structural specificity of 3-chlorolactaldehyde for the sperm isoform. The contraceptive activity of alpha-chlorohydrin and its apparent specificity for the sperm isoform in vivo are likely to be due to differences in metabolism to 3-chlorolactaldehyde in spermatozoa and somatic cells. However, further detailed analysis of the sperm glyceraldehyde-3-phosphate dehydrogenase structure revealed sites in the enzyme that do show significant difference compared with published somatic glyceraldehyde-3-phosphate dehydrogenase structures that could be exploited by structure-based drug design to identify leads for novel male contraceptives.

PubMed: 19542219
DOI: 10.1074/JBC.M109.004648

Experimental method

X-RAY DIFFRACTION (2.38 Å)