Glutamine purine synthesis

The amino acids glycine, aspartate, and glutamine are used in purine synthesis. 

Orotic acid in the diet (usually at a concentration of 1 per cent) can induce a deficiency of adenine and pyridine nucleotides in rat liver (but not in mouse or chick liver). The consequence is to inhibit secretion of lipoprotein into the blood, followed by the depression of plasma lipids, then in the accumulation of triglycerides and cholesterol in the liver (fatty liver) [141 — 161], This effect is not prevented by folic acid, vitamin B12, choline, methionine or inositol [141, 144], but can be prevented or rapidly reversed by the addition of a small amount of adenine to the diets [146, 147, 149, 152, 162]. The action of orotic acid can also be inhibited by calcium lactate in combination with lactose [163]. It was originally believed that the adenine deficiency produced by orotic acid was caused by an inhibition of the reaction of PRPP with glutamine in the de novo purine synthesis, since large amounts of PRPP are utilized for the conversion of orotic acid to uridine-5 -phosphate. However, incorporation studies of glycine-1- C in livers of orotic acid-fed rats revealed that the inhibition is caused rather by a depletion of the PRPP available for reaction with glutamine than by an effect on the condensation itself [160]. 

Figure 10-1. Overview of purine synthesis. Details of the first two reactions and sources of the atoms of the purine ring in inosine 5 -monophosphate (IMP) are shown. PRPP, 5 -phosphoribosyl-1-pyrophosphate Gin, glutamine Gly, glycine Asp, aspartate THF, tetrahydrofolate.

Three major feedback mechanisms cooperate in regulating the overall rate of de novo purine nucleotide synthesis and the relative rates of formation of the two end products, adenylate and guanylate (Fig. 22-35). The first mechanism is exerted on the first reaction that is unique to purine synthesis—transfer of an amino group to PRPP to form 5-phosphoribosylamine. This reaction is catalyzed by the allosteric enzyme glutamine-PRPP amidotransferase, which is inhibited by the end products IMP, AMP, and GMP. AMP and GMP act synergisti-cally in this concerted inhibition. Thus, whenever either AMP or GMP accumulates to excess, the first step in its biosynthesis from PRPP is partially inhibited. 

Many lines of evidence indicate that the first committed step in de novo purine nucleotide biosynthesis, production of 5-phosphoribosylamine by glutamine PRPP amidotransfer-ase, is rate-limiting for the entire sequence. Consequently, regulation of this enzyme is probably the most important factor in control of purine synthesis de novo (fig. 23.24). The enzyme is inhibited by purine-5 -nucleotides, but the most inhibitory nucleotides vary with the source of the enzyme. Inhibition constants (A, ) are usually in the range 10-3-10-5 M. The maximum effect of this end-product inhibition is produced by certain combinations of nucleotides (e.g., AMP and GMP) in optimum concentrations and ratios, indicating two kinds of inhibitor binding sites. This is an example of a concerted feedback inhibition. 

The pathway to purine synthesis is rather long. Parts of the purine molecule are from glycine, tetrahydrofo-late (a one-carbon donor), glutamine, CO and aspartate (fig. 5.4) (B-3). This pathway enters the purine section of the DNA Funhouse at IMP" (Inosine monophosphate), at the ground level. The body can avoid part of the long... 

The committed, regulated step in the pathway catalyzed by PRPP amidotransferase. Note glutamine provides the nitrogen to initiate purine synthesis. 

C. Options A and B are true for purine but not pyrimidine biosynthesis. During pyrimidine synthesis, the entire aspartate molecule is incorporated into the ring. Glutamine is the substrate for carbamoyl phosphate synthetase II, the enzyme involved in pyrimidine biosynthesis. (NH4+ is the substrate for synthetase I used in urea synthesis.) Glydne supplies one nitrogen for purine synthesis. 

The answer is c. (Ivlurray, pp 375— /O I. Scrivt i, pp 2513—2570. Sack, pp 121—138. Wilson, pp 287—320.1 Several control sites exist in the path of purine synthesis where feedback inhibition occurs, AMP, GMP, or IMP may inhibit the first step of the pathway, which is the synthesis ol 5-phosphoribosyl-l-pyrophosphate (PRPP). PRPP synthetase is specifically inhibited. All three nucleotides can inhibit glutamine PRPP aminotranslerase, which catalyzes the second step of the. pathway. AMP blocks the conversion ol IMP to adenylosuccinate. GMP inhibits the lormation ol xanthylate Irom IMP Thus, blockage rather than enhancement ol IMP metabolism to AMP and GMP effectively inhibits purine biosynthesis. 

The substrate for this reaction, ct-D-ribose-5-phosphate, is a product of the pentose phosphate pathway.) Figure 14.24 illustrates the initial phase in the pathway by which PRPP is converted to inosine monophosphate (inosinate), the first purine nucleotide. The process begins with the displacement of the pyrophosphate group of PRPP by the amide nitrogen of glutamine in a reaction catalyzed by glutamine PRPP amidotransferase. This reaction is the committed step in purine synthesis. The product formed is 5-phospho-/3-D-ribosylamine. 

In the next step, which is the first step uniquely related to purine synthesis, the amide nitrogen from glutamine is added to the PRPP to form 5-phosphoribosylamine, catalyzed by PRPP amidotransferase. This step can be inhibited by azaserine, an antimetabolite of glutamine. Glycine is then added, forming an amide bond. This re-... 

The de novo pathway of purine synthesis is complex, consisting of 11 steps, and requiring 6 molecules of ATP for every purine synthesized. The precursors that donate components to produce purine nucleotides include glycine, ribose 5-phosphate, glutamine, aspartate, carbon dioxide, and N -formyl FH4 (Fig. 41.1). Purines are synthesized as ribonucleotides, with the initial purine synthesized being inosine monophosphate (IMP). Adenosine monophosphate (AMP) and guano sine monophosphate (GMP) are each derived from IMP in two-step reaction pathways. 

The committed step of purine synthesis is the formation of 5-phosphoribosyl 1-amine by glutamine phosphoribosyl amidotransferase. This enzyme is strongly inhibited by GMP and AMP (the end products of the purine biosynthetic pathway). The enzyme is also inhibited by the corresponding nucleoside di- and triphosphates, but under cellular conditions, these compounds probably do not play a central role in regulation. The active enzyme is a monomer of 133,000 daltons but is converted to an inactive dimer (270,000 daltons) by binding of the end products. 

Fig. 41.9. The regulation of purine synthesis. PRPP synthetase has two distinct allosteric sites, one for ADP, the other for GDP. Glutamine phosphoribosyl amidotransferase contains adenine nucleotide and guanine nucleotide binding sites the monophosphates are the most important, although the di- and tri-phosphates will also bind to and inhibit the enzyme. Adenylosuccinate synthetase is inhibited by AMP IMP dehydrogenase is inhibited by GMP.

The 5 phosphoribosyl-l-pyrophosphate required for purine synthesis is obtained from ATP and ribose-5-phosphate by reaction (11.39). Glutamine, which is also required, is obtained from glutamic acid by reaction (11.44), and the latter is obtained from a - oxoglutaric acid by reaction (11.124). The last reaction links the Krebs cycle with amino acid metabolism. 

CH-C0-CH2-CH(NH2)-C00H, an antagonist of glutamine. It inhibits de novo purine synthesis in bacteria and mammals. It prevents the growth of experimental tumors, but is toxic for animals. 

Glutamine is present in abundance in some tissues, such as brain, but is scarce in many tumor cells and is a rate-limiting factor in some cells, both in vivo and in vitro it may even be limiting in liver under certain conditions (17, 37). Similarly, glycine and aspartate can be rate-limiting for purine synthesis in some cells (17,38,39). As discussed above, aspartate can some-... 

An increase in the amount of glutamine available for purine biosynthesis could be due to accelerated synthesis, decreased use in other pathways, or decreased breakdown. The third possibility is unlikely, since the activity of glutaminase was found to be normal in a patient with severe hyperuricemia. The possibility of an increase in glutamine synthetase activity cannot be excluded. Inborn errors resulting from increased enzyme activities are rare yet an increase in the activity of (5-aminolevulinic acid occurs in patients with porphyria. All of the metabolic alterations described in gout would satisy a situation in which glutamine use in pathways other than purine synthesis is reduced, but no such alterations have been conclusively demonstrated. 

Both AMP and GMP inhibited purine synthesis at the level of formation of phosphoribosylamine irrespective of whether glutamine or ammonia was the N-donor. Detailed analysis of the AMP studies however was difficult because of the rapid enzymatic deamination of AMP with this enzyme preparation in the absence of GTP. 

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