Stereochemistry of biosynthesis of the vinyl groups of protoporphyrin-IX: a short synthesis of porphobilinogen

Battersby, Alan R.; McDonald, Edward; Wurziger, H.; James, Kevin J.

doi:10.1039/c39750000493

Cited by 35 publications

(21 citation statements)

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“…Porphobilinogen was either synthesised enzymically using purified 5-aminolaevulinic acid dehydratase (Jordan and Seehra 1986) or chemically (Battersby et al 1975) in which case it was a gift from Dr. C. Abell, Department of Chemistry, University of Cambridge, UK. Dalton VII molecular-weight standards, Sephadex G-75, diethylaminoethyl (DEAE) Sephacel, Freund's complete and incomplete adjuvant, protein A-Sepharose and general laboratory chemicals were purchased from Sigma Chemical Co, Poole, Dorset, UK.…”

Section: Methodsmentioning

confidence: 99%

Subcellular location of the tetrapyrrole synthesis enzyme porphobilinogen deaminase in higher plants: an immunological investigation

Witty

Jones

Robb

et al. 1996

Planta

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A recombinant plasmid, pArab8, harbouring the cDNA encoding the mature form of the tetrapyrrole synthesis enzyme porphobilinogen deaminase (EC 4.3.1.8; also known as hydroxymethylbilane synthase) from Arabidopsis thaliana (L.) Heynh. has been constructed, and used to transform Escherichia coli. The porphobilinogen deaminase protein from Arabidopsis was overexpressed in this strain, and purified to homogeneity (3000-fold) with a yield of 20%. Antibodies were raised against the purified plant enzyme, and used in Western blot analysis, immunoprecipitation of enzyme activity and immuno-gold electron microscopy. The results indicate that the enzyme is confined to plastids in both leaves and roots. The implications of this finding for plant tetrapyrrole synthesis are discussed.

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Section: Methodsmentioning

confidence: 99%

Subcellular location of the tetrapyrrole synthesis enzyme porphobilinogen deaminase in higher plants: an immunological investigation

Witty

Jones

Robb

et al. 1996

Planta

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“…The exocyclic double bond is formed by proton abstraction at the ␤-position of the substrate propionate side chain by peroxide and the resulting six-membered-ring transition. The reaction concludes with the elimination of CO 2 and H 2 O 2 followed by bond rearrangements with the formation of the product vinyl group (160,(245)(246)(247)(248). Currently, the structures of CgdC from yeast (PDB accession number 1TLB), human (accession number 2AEX), Leishmania major (accession number 3DWR), Leishmania donovani (accession number 3EJO), Leishmania naiffi (accession number 3E8J), and Acinetobacter baumannii (accession number 5EO6) have been solved, with all of them revealing an unprecedented fold for the monomer of large seven-stranded antiparallel ␤-sheets covered on both sides by ␣-helices (Fig.…”

Section: Conversion Of Coproporphyrinogen III Into Protoporphyrinogenmentioning

confidence: 99%

Prokaryotic Heme Biosynthesis: Multiple Pathways to a Common Essential Product

Dailey

Gerdes³

et al. 2017

Microbiol Mol Biol Rev

258

335

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SUMMARY The advent of heme during evolution allowed organisms possessing this compound to safely and efficiently carry out a variety of chemical reactions that otherwise were difficult or impossible. While it was long assumed that a single heme biosynthetic pathway existed in nature, over the past decade, it has become clear that there are three distinct pathways among prokaryotes, although all three pathways utilize a common initial core of three enzymes to produce the intermediate uroporphyrinogen III. The most ancient pathway and the only one found in the Archaea converts siroheme to protoheme via an oxygen-independent four-enzyme-step process. Bacteria utilize the initial core pathway but then add one additional common step to produce coproporphyrinogen III. Following this step, Gram-positive organisms oxidize coproporphyrinogen III to coproporphyrin III, insert iron to make coproheme, and finally decarboxylate coproheme to protoheme, whereas Gram-negative bacteria first decarboxylate coproporphyrinogen III to protoporphyrinogen IX and then oxidize this to protoporphyrin IX prior to metal insertion to make protoheme. In order to adapt to oxygen-deficient conditions, two steps in the bacterial pathways have multiple forms to accommodate oxidative reactions in an anaerobic environment. The regulation of these pathways reflects the diversity of bacterial metabolism. This diversity, along with the late recognition that three pathways exist, has significantly slowed advances in this field such that no single organism's heme synthesis pathway regulation is currently completely characterized.

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“…Bacto tryptone and Bacto yeast extract were obtained from Difco Laboratories. Porphobilinogen was synthesized as previously described [20]. All other chemicals were of analytical reagent grade and were obtained from Merck or from Fluka.…”

Section: Generalmentioning

confidence: 99%

Purification, characterization, crystallisation and X‐ray analysis of selenomethionine‐labelled hydroxymethylbilane synthase from Escherichia coli

Hädener

Matzinger

Malashkevich

et al. 1993

European Journal of Biochemistry

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Hydroxymethylbilane synthase (HMBS) catalyses the conversion of porphobilinogen into hydroxymethylbilane, a linear tetrapyrrolic intermediate in the biosynthesis of haems, chlorophylls, vitamin B,, and related macrocycles. In the course of an investigation of the crystal structure of this enzyme, we intended to follow a new strategy to obtain the X-ray phase information, i. e. the collection of multiwavelength anomalous diffraction data from a crystal of a seleno-L-methionine (SeMet)-labelled variant of the protein. We have expressed and purified HMBS from Escherichia coli (34268 Da) in which all (six) methionine (Met) residues are replaced by SeMet. Complete replacement, as shown by amino acid composition analysis and by electrospray mass spectrometry, was achieved by growing the Met-requiring mutant E. coli PO1562 carrying the plasmid pPA410 in a medium containing 50mg/l SeMet as the sole source of Met. [SeMetIHMBS exhibits full enzyme activity, as reflected by unchanged steady-state kinetic parameters relative to native enzyme. Rhombohedral crystals of [SeMetIHMBS could be grown at the pH optimum (7.4) of the enzyme (solutions containing 30 mg/ml protein, 0.4 mh4 EDTA, 20 mM dithiothreitol, 3 M NaCl and 15 mM Bistris-propane buffer were equilibrated by vapour diffusion at 20 "C against reservoirs of saturated NaC1). However, being very thin plates, these crystals were not suitable for X-ray analysis. Alternatively, rectangular crystals were obtained at pH 5.3 using conditions based on those reported for wild-type HMBS [sitting drops of 50 p1 containing 6-7 mg/ml protein, 0.3 mh4 EDTA, 15 mM dithiothreitol, 10 % (mass/vol.) poly(ethy1ene glycol) 6000 and 0.01 % NaN, in 0.1 M sodium acetate were equilibrated by vapour diffusion at 20°C against a reservoir of 10-20 mg solid dithiothreitol]. X-ray diffraction data of the crystals were complete to 93.8% at 0.21 nm resolution and showed that [SeMetIHMBS and native HMBS crystallise isomorphously. A difference Fourier map using FseMet -Fnative and phases derived from the native structure, which has recently been determined independently by multiple isomorphous replacement, showed positive difference peaks centered at or close to where the sulphur atoms of the Met side chains appear in the native structure. In addition, paired positivehegative peaks in the difference map near the cofactor of HMBS indicate conformational differences in the active site, probably due to differences in the state of oxidation of the cofactor in the two crystalline samples.The replacement of methionine residues by seleno-Lmethionine (SeMet) in proteins is part of a new strategy to tackle the cardinal problem of protein crystallography : the determination of phases [l]. Given the availability of welldiffracting crystals of a SeMet-containing protein, the ap- proach involves the collection of multiwavelength anomalous diffraction (MAD) data from a single crystal using synchrotron radiation. Specific algebraic methods for MAD data analysis allow the evaluation of phase angles for the diffracte...

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Stereochemistry of biosynthesis of the vinyl groups of protoporphyrin-IX: a short synthesis of porphobilinogen

Abstract: SummavyPorphobilinogen (1) , [aH]-la%eXed in the propionic residue, is synthesised by a short route and is used to establish that both vinyl groups of protoporphyrin-IX are biosynthesised by overall antiperiplanar elimination of a proton and carbon dioxide. 731 and refs. therein.

Cited by 35 publications

References 0 publications

Subcellular location of the tetrapyrrole synthesis enzyme porphobilinogen deaminase in higher plants: an immunological investigation

Subcellular location of the tetrapyrrole synthesis enzyme porphobilinogen deaminase in higher plants: an immunological investigation

Prokaryotic Heme Biosynthesis: Multiple Pathways to a Common Essential Product

Purification, characterization, crystallisation and X‐ray analysis of selenomethionine‐labelled hydroxymethylbilane synthase from Escherichia coli

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