Porphyrin biosynthesis, control

The tetramerization of suitable monopyrroles is one of the simplest and most effective approaches to prepare porphyrins (see Section 1.1.1.1.). This approach, which is best carried out with a-(hydroxymethyl)- or ot-(aminomethyl)pyrroles, can be designated as a biomimetic synthesis because nature also uses the x-(aminomethyl)pyrrole porphobilinogen to produce uroporphyrinogen III. the key intermediate in the biosynthesis of all kinds of naturally occurring porphyrins, hydroporphyrins and corrins. The only restriction of this tetramerization method is the fact that tnonopyrroles with different -substituents form a mixture of four constitutionally isomeric porphyrins named as porphyrins I, II, III, and IV. In the porphyrin biosynthesis starting from porphobilinogen, which has an acetic acid and a propionic acid side chain in the y6-positions, this tetramerization is enzymatically controlled so that only the type III constitutional isomer is formed. 

Formation of S-aminolevulinic acid (ALA) All the carbon and nitrogen atoms of the porphyrin molecule are provided by two simple building blocks glycine (a nonessential amino acid) and succinyl CoA (an intermediate in the citric acid cycle). Glycine and succinyl CoA condense to form ALA in a reaction catalyzed by ALA synthase (Figure 21.3) This reaction requires pyridoxal phosphate as a coenzyme, and is the rate-controlling step in hepatic porphyrin biosynthesis. 

One obvious place to control porphyrin biosynthesis is to control the synthesis of 5-AL. This is the only step which requires a high enei bond in the form of succinyl-CoA. All the other steps involve reactions which are largely irrevermble and thermodynamically favored, such as the formation of a pyrrole ring, decarboxylation, and oxidation to the aromatic porphyrin ring. The rate of synthesis of 5-AL may be controlled by the amount of the enzyme 5-AL-f thetase, by the concentration of its coenzyme-pyridoxal phosphate, by the steady state level of succinyl-CoA and of glycine, and possibly by inhibitors of the enzyme such as cysteine. Once formed the 5-AL may be converted to porphyrin or may be oxidized via the Shemin cycle. It is difficult to obtain a quantitative estimate of the importance of this oxidative pathway (65, S91). 

The compartmentation of 5-AL-synthetase inside the mitochondrion and 5-AL-dehydrase outside suggests still another mechanism of control of porphyrin biosynthesis, namely a permeability control for 6-AL at the membrane as indicated in figure 31. This figure is also a summary of the known steps of porphyrin biosynthesis, of their tentatively assigned locations in the cell, and of possible sites of enzymic change in the porphyria diseases. 

Burnham, B. F., and Lascelles, J. (1963). Control of porphyrin biosynthesis through a negative-feedback mechanism. Biochem. J. 87, 462-472. 

The similarity between the structures of the corrinoids and the porphyrins becomes evident from comparison of cobyrinic acid (75) (the simplest of the corronoids so far isolated) with uroporphyrinogen III (70). The possibility of a biosynthetic relationship between these structures was suggested by Shemin, who reported the incorporation of [14C]ALA into vitamin Bn and confirmed by the subsequent demonstration that PBG was also incorporated. The ubiquitous precursorial role of uroporphyrinogen III in heme, chlorophyll and corrinoid biosynthesis proposed by Porra (65BBA(107)176) was, however, not substantiated by experimental evidence until much later, when under carefully controlled conditions cells of Propionibacterium shermanii were shown to incorporate radioactivity from [14C]uroporphyrinogen III into vitamin Bn (72JA8269). 

We devised a screen for isolating mutants defective in iron-dependent regulation of heme biosynthesis that did not require prior knowledge of the mechanism or of the rate-limiting steps [83]. We speculated that if the pathway as a whole were regulated by iron, a mutant defective in that control would accumulate protoporphyrin under iron limitation. Mutants defective in the heme synthesis enzymes ferrochelatase [75] or protoporphyrinogen oxidase would likely have a similar phenotype, but porphyrin accumulation would likely be independent of iron in the structural gene mutants, and those strains would also be expected to be heme auxotrophs. 

The Chi synthesis modulators that Rebeiz et al. (13, 14) used in conjunction with ALA could be divided into three categories A) enhancers of ALA conversion to porphyrins (2-pyridine aldoxime, 2-pyridine aldehyde, picolinic acid, 2,2 dipyridyl disulfide, 2,2 -dipyridyl amine, 4,4 dipyridyl, and phenanthridine), B) inducers of ALA biosynthesis and porphyrin accumulation (2,2 -dipyridyl and 1,10-phenanthroline), and C) inhibitors of MV PChlide synthesis (2,3-dipyridyl, 2,4-dipyridyl, 1,7-phenanthroline, and 4,7-phenanthroline). Compounds in group A did not cause significant porphyrin accumulation alone however, they enhanced dark conversion of exogenous ALA to porphyrins. This group was further subdivided into compounds that enhanced conversion of ALA to MV PChlide (2-pyridine aldoxime, 2-pyridine aldehyde, picolinic acid, and 2,2 -dipyridyl disulfide) and those that stimulated conversion to DV PChlide (4,4 dipyridyl, 2,2 dipyridyl amine, and phenanthridine). To qualify as an ALA biosynthesis and porphyrin accumulation inducer (category B), the compound had to cause these effects in the absence of ALA. Compounds in category C had to inhibit accumulation of MV PChlide with or without ALA. In most cases, in conjunction with ALA, the compounds stimulated DV PChlide accumulation compared to the ALA-treated control. 

Granick, S., and Kappas, A. (1967). Steroid control of porphyrin and heme biosynthesis A new biological function of steroid hormone metabolites. Proc. Natl. Acad. Sci. U.S. 57, 1463-1467. 

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