Matches in SemOpenAlex for { <https://semopenalex.org/work/W4237093225> ?p ?o ?g. }
Showing items 1 to 56 of
56
with 100 items per page.
- W4237093225 abstract "[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT REQUEST OF AUTHOR.] Pyrroline-5-carboxylate reductase (PYCR) is the final enzyme in proline biosynthesis, catalyzing the NAD(P)H-dependent reduction of [Delta]1-pyrroline-5-carboxylate (P5C) to proline. Mutations in the PYCR1 gene alter mitochondrial function and cause the connective tissue disorder cutis laxa. Furthermore, PYCR1 is overexpressed in multiple cancers, and the PYCR1 knockout suppresses tumorigenic growth, suggesting PYCR1 is a potential cancer target. However, inhibitor development has been stymied by limited mechanistic details for the enzyme, particularly in light of a previous crystallographic study that placed the cofactor binding site in the C-terminal domain rather than the anticipated Rossmann fold of the N-terminal domain. To fill this gap, we report crystallographic, sedimentation velocity, and kinetics data for human PYCR1. Structures of binary complexes of PYCR1 with NADPH or proline determined at 1.9 A resolution provide insight into cofactor and substrate recognition. We see NADPH bound to the Rossmann fold, over 25 A from the previously proposed site. The 1.85 A resolution structure of a ternary complex containing NADPH and a P5C/proline analog provides a model of the Michaelis complex formed during hydride transfer. Sedimentation velocity shows that PYCR1 forms a concentration-dependent decamer in solution, consistent with the pentamer-of-dimers assembly seen crystallographically. Kinetic and mutational analysis confirmed several features seen in the crystal structure, including the importance of a hydrogen bond between Thr238 and the substrate as well as limited cofactor discrimination. We also report kinetic and structural data for PYCR1 complexed with multiple P5C/Pro analogs to probe the potential of PYCR1 as a cancer therapy target. Crystal structures of binary complexes of PYCR1 with L-tetrahydro-2-furoic acid (THFA),N-formyl L-proline (NFLP), thiazolidine-2-carboxylate (T2C), and thiazolidine-4-carboxylate (T4C) have been determined at 1.80-2.35 A resolution. We also present inhibition data for the forward reaction of P5C reduction to proline in the presence of each analog." @default.
- W4237093225 created "2022-05-12" @default.
- W4237093225 creator A5021840577 @default.
- W4237093225 date "2021-04-14" @default.
- W4237093225 modified "2023-10-17" @default.
- W4237093225 title "Structural, biochemical, and inhibition studies of proline biosynthetic enzymes" @default.
- W4237093225 doi "https://doi.org/10.32469/10355/76143" @default.
- W4237093225 hasPublicationYear "2021" @default.
- W4237093225 type Work @default.
- W4237093225 citedByCount "0" @default.
- W4237093225 crossrefType "dissertation" @default.
- W4237093225 hasAuthorship W4237093225A5021840577 @default.
- W4237093225 hasConcept C181199279 @default.
- W4237093225 hasConcept C185592680 @default.
- W4237093225 hasConcept C18903297 @default.
- W4237093225 hasConcept C197957613 @default.
- W4237093225 hasConcept C2777289219 @default.
- W4237093225 hasConcept C2778074193 @default.
- W4237093225 hasConcept C2779815552 @default.
- W4237093225 hasConcept C41183919 @default.
- W4237093225 hasConcept C515207424 @default.
- W4237093225 hasConcept C55493867 @default.
- W4237093225 hasConcept C71240020 @default.
- W4237093225 hasConcept C8010536 @default.
- W4237093225 hasConcept C86756495 @default.
- W4237093225 hasConcept C86803240 @default.
- W4237093225 hasConceptScore W4237093225C181199279 @default.
- W4237093225 hasConceptScore W4237093225C185592680 @default.
- W4237093225 hasConceptScore W4237093225C18903297 @default.
- W4237093225 hasConceptScore W4237093225C197957613 @default.
- W4237093225 hasConceptScore W4237093225C2777289219 @default.
- W4237093225 hasConceptScore W4237093225C2778074193 @default.
- W4237093225 hasConceptScore W4237093225C2779815552 @default.
- W4237093225 hasConceptScore W4237093225C41183919 @default.
- W4237093225 hasConceptScore W4237093225C515207424 @default.
- W4237093225 hasConceptScore W4237093225C55493867 @default.
- W4237093225 hasConceptScore W4237093225C71240020 @default.
- W4237093225 hasConceptScore W4237093225C8010536 @default.
- W4237093225 hasConceptScore W4237093225C86756495 @default.
- W4237093225 hasConceptScore W4237093225C86803240 @default.
- W4237093225 hasLocation W42370932251 @default.
- W4237093225 hasOpenAccess W4237093225 @default.
- W4237093225 hasPrimaryLocation W42370932251 @default.
- W4237093225 hasRelatedWork W137495 @default.
- W4237093225 hasRelatedWork W15085303 @default.
- W4237093225 hasRelatedWork W15160677 @default.
- W4237093225 hasRelatedWork W19015500 @default.
- W4237093225 hasRelatedWork W19495342 @default.
- W4237093225 hasRelatedWork W36952825 @default.
- W4237093225 hasRelatedWork W37554847 @default.
- W4237093225 hasRelatedWork W8834194 @default.
- W4237093225 hasRelatedWork W9097482 @default.
- W4237093225 hasRelatedWork W15884920 @default.
- W4237093225 isParatext "false" @default.
- W4237093225 isRetracted "false" @default.
- W4237093225 workType "dissertation" @default.