title: New insights into the mechanism of substrates trafficking in Glyoxylate/Hydroxypyruvate reductases.
abstract: Glyoxylate accumulation within cells is highly toxic. In humans, it is associated with hyperoxaluria type 2 (PH2) leading to renal failure. The glyoxylate content within cells is regulated by the NADPH/NADH dependent glyoxylate/hydroxypyruvate reductases (GRHPR). These are highly conserved enzymes with a dual activity as they are able to reduce glyoxylate to glycolate and to convert hydroxypyruvate into D-glycerate. Despite the determination of high-resolution X-ray structures, the substrate recognition mode of this class of enzymes remains unclear. We determined the structure at 2.0 Å resolution of a thermostable GRHPR from Archaea as a ternary complex in the presence of D-glycerate and NADPH. This shows a binding mode conserved between human and archeal enzymes. We also determined the first structure of GRHPR in presence of glyoxylate at 1.40 Å resolution. This revealed the pivotal role of Leu53 and Trp138 in substrate trafficking. These residues act as gatekeepers at the entrance of a tunnel connecting the active site to protein surface. Taken together, these results allowed us to propose a general model for GRHPR mode of action.
authors: Univ. Grenoble Alpes, IBS, F-38044 Grenoble, France.|CNRS, IBS, F-38044 Grenoble, France.|CEA, IBS, F-38044 Grenoble, France.
authors: Louise
authors: L
authors: Lassalle
authors: Univ. Grenoble Alpes, IBS, F-38044 Grenoble, France.|CNRS, IBS, F-38044 Grenoble, France.|CEA, IBS, F-38044 Grenoble, France.
authors: Sylvain
authors: S
authors: Engilberge
authors: Univ. Grenoble Alpes, IBS, F-38044 Grenoble, France.|CNRS, IBS, F-38044 Grenoble, France.|CEA, IBS, F-38044 Grenoble, France.
authors: Dominique
authors: D
authors: Madern
authors: Univ. Grenoble Alpes, IBS, F-38044 Grenoble, France.|CNRS, IBS, F-38044 Grenoble, France.|CEA, IBS, F-38044 Grenoble, France.
authors: Pierre
authors: P
authors: Vauclare
authors: Univ. Grenoble Alpes, IBS, F-38044 Grenoble, France.|CNRS, IBS, F-38044 Grenoble, France.|CEA, IBS, F-38044 Grenoble, France.
authors: Bruno
authors: B
authors: Franzetti
authors: Univ. Grenoble Alpes, IBS, F-38044 Grenoble, France.|CNRS, IBS, F-38044 Grenoble, France.|CEA, IBS, F-38044 Grenoble, France.
authors: Eric
authors: E
authors: Girard
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D057075:Enzyme Assays; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005988:Glyceric Acids / Q000737:chemistry / Q000378:metabolism; D006038:Glyoxylates / Q000737:chemistry / Q000378:metabolism; D050537:Hydroxypyruvate Reductase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D009243:NAD / Q000737:chemistry / Q000378:metabolism; D009249:NADP / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D055550:Protein Stability; D019711:Pyrococcus / Q000737:chemistry* / Q000201:enzymology; D020027:Pyrococcus furiosus / Q000737:chemistry* / Q000201:enzymology; D043902:Pyrococcus horikoshii / Q000737:chemistry* / Q000201:enzymology; D011773:Pyruvates / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D019843:Archaeal Proteins; D005988:Glyceric Acids; D006038:Glyoxylates; D011773:Pyruvates; D011994:Recombinant Proteins; D009243:NAD; D009249:NADP; C042971:glyceric acid; C012375:hydroxypyruvic acid; D000429:Alcohol Oxidoreductases; C020830:glyoxylate reductase; D050537:Hydroxypyruvate Reductase; C031150:glyoxylic acid