Spectroscopic evidence for a porphobilinogen deaminase-tetrapyrrole complex that is an intermediate in the biosynthesis of uroporphyrinogen III.

  title={Spectroscopic evidence for a porphobilinogen deaminase-tetrapyrrole complex that is an intermediate in the biosynthesis of uroporphyrinogen III.},
  author={S Ros{\'e} and ROSALIA B. Frydman and Carlos R. de los Santos and Adriana R. Sburlati and Aldonia Valasinas and Benjamin J Frydman},
  volume={27 13},
Incubation of porphobilinogen (PBG) with PBG deaminase from Rhodopseudomonas sphaeroides in carbonate buffer (pH 9.2) to total PBG consumption resulted in low yields of uroporphyrinogen I (uro'gen I). In the reaction mixture a pyrrylmethane accumulated, which at longer incubation periods was transformed into uro'gen I. The accumulated pyrrylmethane gave an Ehrlich reaction which was different from that of a 2-(aminomethyl)dipyrrylmethane or 2-(aminomethyl)tripyrrane. It resembled that of a… Expand
Purification of Porphobilinogen Deaminase and the Sequencing and Expression of thehemC Gene of Rhodobacter Capsulatus.
Rhodobacter capsulatus uses the common tetrapyrrole pathway to synthesize heme, bacteriochlorophyll, siroheme and vitamin B-12. As part of our studies on the regulation of this pathway, the productExpand
Site-directed mutagenesis and high-resolution NMR spectroscopy of the active site of porphobilinogen deaminase.
The NMR spectrum of the covalent complex of deaminase with the suicide inhibitor 2-bromo-[2,11-13C2]PBG reveals that the aninomethyl terminus of the inhibitor reacts with the enzyme's cofactor at the alpha-free pyrrole. Expand
Purification and kinetic studies on a porphobilinogen deaminase from the unicellular green alga Scenedesmus obliquus
Porphobilinogen deaminase, the enzyme condensing four molecules of porphobilinogen, was isolated and purified from light grown Scenedesmus obliquus (wild type). The purification procedure includedExpand
Biosynthesis of Hemes.
  • S. Beale
  • Biology, Medicine
  • EcoSal Plus
  • 2007
Several lines of evidence converge to support a regulatory model in which the cellular level of available or free protoheme controls the rate of heme synthesis at the level of the first step unique to heme synthesisation, the formation of GSA by the action of GTR. Expand
Biosynthesis and Structures of Porphyrins and Hemes
Anoxygenic phototrophs are capable of synthesizing all three major types of tetrapyrroles: hemes, chlorophylls, and corrins. These molecules are derived from a common, branched biosynthetic pathway.Expand
Biosynthesis of Chlorophylls and Hemes
Publisher Summary This chapter focuses on tetrapyrrole metabolism as a general process that occurs in all chloroplast-containing organisms, but stresses results that have been obtained withExpand
NMR Studies of the Enzyme Mechanisms of Vitamin B12 Biosynthesis
For the past 20 years our laboratory has been engaged in the elucidation of the vitamin B12 biosynthetic pathway.1. By 1977 it had been established that 5-aminolevulinic acid (ALA), porphobilinogenExpand
Isolation and Characterization of a Rhodobacter Capsulatus hemC Mutant.
Rhodobacter capsulatus, a purple, nonsulfur photosynthetic bacterium, uses the tetrapyrrole pathway to synthesize four end products: heme, bacteriochlorophyll, vitamin B12, and siroheme. ThisExpand
Mechanistic and evolutionary aspects of vitamin B12 biosynthesis
For the past 20 years our laboratory has been engaged in the elucidation of the vitamin B12 biosynthetic pathway and a previous Account1 described progress made in the first ten years of thisExpand
Biosynthesis of Cyanobacterial Tetrapyrrole Pigments: Hemes, Chlorophylls, and Phycobilins
Cyanobacteria are versatile tetrapyrrole synthesizers that are able to produce end products representing all major branches of the tetrapyrrole biosynthetic pathway: hemes, chlorophylls, phycobilins,Expand


Biosynthesis of uroporphyrinogens. Interaction among 2-(aminomethyl)bilanes and the enzymatic system.
An outline of the most likely pathway of uroporphyrinogen III biosynthesis from porphobilinogen is given and results obtained with 2-aminomethyldipyrrylmethanes are obtained. Expand
Biosynthesis of porphyrins and corrins. 2. Isolation, purification, and NMR investigations of the porphobilinogen-deaminase covalent complex.
During the resultant formation of [3H]uro'gen I, a transient low-intensity signal was detected that has been tentatively assigned to the highly reactive azafulvene species, proposed in several mechanistic schemes for porphyrin biosynthesis. Expand
The isolation and characterization of catalytically competent porphobilinogen deaminase—intermediate complexes
It is established that the enzyme forms a relatively stable covalent link with the bound pyrrole residues and the appearance of 3-intermediate complexes in the case of the bacterial deaminase suggests that discrete enzyme intermediate complexes exist. Expand
Biosynthesis of porphyrins and related macrocycles. Part 21. The interaction of deaminase and its product (hydroxymethylbilane) and the relationship between deaminase and cosynthetase
Experiments are described which show that (a) deaminase is inhibited by its own product, the linear hydroxymethylbilane; (b) this inhibition of uptake of porphobilinogen (PBG) is competitive,Expand
Biosynthesis of uroporphyrinogens from porphobilinogen: mechanism and the nature of the process.
Different enzyme-bound dipyrrylmethanes are formed form the beginning of the process, and this can be demonstrated by using synthetic dipyrries and tripyrranes. Expand
Biosynthesis of porphyries and related macrocycles. Part 17. Chemical and enzymic transformation of isomeric aminomethylbilanes into uroporphyrinogens: proof that unrearranged bilane is the preferred enzymic substrate and detection of a transient intermediate
Six isomeric aminomethylbilanes have been built by unambiguous synthesis. One bilane has the unrearranged structure corresponding to straightforward head-to-tail assembly of four units ofExpand
Chemistry of porphyrin biosynthesis: results and applications.
Systematic investigation of the in vitro condensation of porphobilinogen derivatives under conditions of kinetic control revealed that the ratio of uroporphyrinogen I in the reaction mixture dropped steadily in favour of the III/IV isomers with increasing acidity. Expand
Biosynthesis of porphyrins and related macrocycles. Part 16. Proof that the single intramolecular rearrangement leading to natural porphyrins (type-III) occurs at the tetrapyrrole level
The unrearranged aminomethylbilane (2) is synthesised by a rational route and is proved to be converted by the enzymes deaminase and cosynthetase, working co-operatively, into uro'gen-III (3). TheExpand
13C NMR studies of porphobilinogen synthase: observation of intermediates bound to a 280,000-dalton protein.
13C NMR has been used to observe the equilibrium complex of [4-13C]ALA bound to porphobilinogen (PBG) synthase (5-aminolevulinate dehydratase), a 280,000-dalton protein, indicating that the predominant species is probably a distorted form of PBG. Expand
The synthesis of porphyrins and bacteriochlorophyll by cell suspensions of Rhodopseudomonas spheroides.
Stich & Eisgruber (1951) found that the amount of coproporphyrin Type I in cells of Saccharomyces anamensis is increased 5to 10-fold by addition of pantothenate to the growth medium, while addition of riboflavin completely suppresses porphyr in formation. Expand