T1 - Regulation of proline biosynthesis
Pyrroline-5-carboxylate reductase (PYCR) is the final enzyme in proline biosynthesis, catalyzing the NAD(P)H-dependent reduction of Δ1-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 knock-out suppresses tumorigenic growth, suggesting that 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 Å resolution provide insight into cofactor and substrate recognition. We see NADPH bound to the Rossmann fold, over 25 Å from the previously proposed site. The 1.85 Å 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 Thr-238 and the substrate as well as limited cofactor discrimination.
MetaCyc L-proline biosynthesis III
is the final enzyme in proline biosynthesis…
Histidine is special in that its biosynthesis is inherently linked to thepathways of nucleotide formation. Histidine residues are often found in enzymeactive sites, where the chemistry of the imidazole ring of histidine makes it anucleophile and a good acid/base catalyzer. We now know that RNA can havecatalytic properties, and there has been speculation that life was originallyRNA-based. Perhaps the transition to protein catalysis from RNA catalysisoccurred at the origin of histidine biosynthesis.
The proline biosynthesis in living organisms | SpringerLink
We will look at this pathway in a bit more detail, because it involves themolecule 5-phosphoribosyl--pyrophosphate (which wewill refer to as "PRPP" from now on). PRPP is also involved in thesynthesis of purines and pyrimidines, as we will soon see. In the first step ofhistidine synthesis, PRPP condenses with ATP to form a purine, N1-5'-phosphoribosylATP, in a reaction that is driven by the subsequent hydrolysis of thepyrophosphate that condenses out. Glutamine again plays a role as an amino groupdonor, this time resulting in the formation of 5-aminoamidazole-4-carboximideribonucleotide (ACAIR), which is an intermediate in purine biosynthesis.