Evidence for a Covalent Intermediate in the S-adenosyl-L-methionine-dependent Transmethylation Reaction Catalysed by Sirohaem Synthase

Overview

Journal Biochem J

Specialty Biochemistry

Date 1996 Jan 15

PMID 8573073

Citations 3

Authors

S C Woodcock

M J Warren

Affiliations

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Abstract

CysG, also known as uroporphyrinogen III methylase and sirohaem synthase (CysG; EC 2.1.1.107), is a multifunctional enzyme that is able to transform uroporphyrinogen III into sirohaem via two S-adenosyl-L-methionine (AdoMet)-dependent transmethylations, an NAD(+)-dependent dehydrogenation and a ferrochelation. The apparent tight binding of AdoMet to this multifunctional enzyme is investigated. The use of a rapid AdoMet binding assay demonstrates that CysG becomes labelled with both [methyl-3H]AdoMet and [carboxyl-14C]AdoMet. Further experiments show that the CysG-AdoMet complex is subsequently able to methylate uroporphyrinogen III. CysG remains associated with the labelled constituents of the AdoMet even after denaturation with urea and SDS/PAGE, suggesting that the AdoMet has become covalently linked to the protein. A rapid examination of some of the other transmethylases involved in corrin biosynthesis reveals that they bind the AdoMet in a similar fashion. A multistep transmethylation mechanism is proposed to explain the observed results.

Citing Articles

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Effect of mutations in the transmethylase and dehydrogenase/chelatase domains of sirohaem synthase (CysG) on sirohaem and cobalamin biosynthesis.

Woodcock S, Raux E, Levillayer F, Thermes C, Rambach A, Warren M Biochem J. 1998; 330 ( Pt 1):121-9.

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References

BATTERSBY A . How nature builds the pigments of life: the conquest of vitamin B12. Science. 1994; 264(5165):1551-7. DOI: 10.1126/science.8202709. View

Satoh S, Yang S . Specificity of S-adenosyl-L-methionine in the inactivation and the labeling of 1-aminocyclopropane-1-carboxylate synthase isolated from tomato fruits. Arch Biochem Biophys. 1989; 271(1):107-12. DOI: 10.1016/0003-9861(89)90260-9. View

Mascaro Jr L, Horhammer R, Eisenstein S, Seller L, Mascaro K, Floss H . Synthesis of methionine carrying a chiral methyl group and its use in determining the steric course of the enzymatic C-methylation of indolepyruvate during indolmycin biosynthesis. J Am Chem Soc. 1977; 99(1):273-4. DOI: 10.1021/ja00443a062. View

Cole J, Newman B, White P . Biochemical and genetic characterization of nirB mutants of Escherichia coli K 12 pleiotropically defective in nitrite and sulphite reduction. J Gen Microbiol. 1980; 120(2):475-83. DOI: 10.1099/00221287-120-2-475. View

Blanche F, Debussche L, THIBAUT D, Crouzet J, Cameron B . Purification and characterization of S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase from Pseudomonas denitrificans. J Bacteriol. 1989; 171(8):4222-31. PMC: 210194. DOI: 10.1128/jb.171.8.4222-4231.1989. View

Warren M, Roessner C, Santander P, Scott A . The Escherichia coli cysG gene encodes S-adenosylmethionine-dependent uroporphyrinogen III methylase. Biochem J. 1990; 265(3):725-9. PMC: 1133693. DOI: 10.1042/bj2650725. View

Warren M, Stolowich N, Santander P, Roessner C, Sowa B, Scott A . Enzymatic synthesis of dihydrosirohydrochlorin (precorrin-2) and of a novel pyrrocorphin by uroporphyrinogen III methylase. FEBS Lett. 1990; 261(1):76-80. DOI: 10.1016/0014-5793(90)80640-5. View

Som S, Friedman S . Direct photolabeling of the EcoRII methyltransferase with S-adenosyl-L-methionine. J Biol Chem. 1990; 265(8):4278-83. View

Peakman T, Crouzet J, Mayaux J, Busby S, Mohan S, Harborne N . Nucleotide sequence, organisation and structural analysis of the products of genes in the nirB-cysG region of the Escherichia coli K-12 chromosome. Eur J Biochem. 1990; 191(2):315-23. DOI: 10.1111/j.1432-1033.1990.tb19125.x. View

10.

Crouzet J, Cauchois L, Blanche F, Debussche L, THIBAUT D, Rouyez M . Nucleotide sequence of a Pseudomonas denitrificans 5.4-kilobase DNA fragment containing five cob genes and identification of structural genes encoding S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase and cobyrinic acid a,c-diamide.... J Bacteriol. 1990; 172(10):5968-79. PMC: 526919. DOI: 10.1128/jb.172.10.5968-5979.1990. View

11.

Crouzet J, Cameron B, Cauchois L, Rigault S, Rouyez M, Blanche F . Genetic and sequence analysis of an 8.7-kilobase Pseudomonas denitrificans fragment carrying eight genes involved in transformation of precorrin-2 to cobyrinic acid. J Bacteriol. 1990; 172(10):5980-90. PMC: 526920. DOI: 10.1128/jb.172.10.5980-5990.1990. View

12.

Wu J, Siegel L, KREDICH N . High-level expression of Escherichia coli NADPH-sulfite reductase: requirement for a cloned cysG plasmid to overcome limiting siroheme cofactor. J Bacteriol. 1991; 173(1):325-33. PMC: 207191. DOI: 10.1128/jb.173.1.325-333.1991. View

13.

Robin C, Blanche F, Cauchois L, Cameron B, Couder M, Crouzet J . Primary structure, expression in Escherichia coli, and properties of S-adenosyl-L-methionine:uroporphyrinogen III methyltransferase from Bacillus megaterium. J Bacteriol. 1991; 173(15):4893-6. PMC: 208169. DOI: 10.1128/jb.173.15.4893-4896.1991. View

14.

Warren M, Roessner C, Ozaki S, Stolowich N, Santander P, Scott A . Enzymatic synthesis and structure of precorrin-3, a trimethyldipyrrocorphin intermediate in vitamin B12 biosynthesis. Biochemistry. 1992; 31(2):603-9. DOI: 10.1021/bi00117a043. View

15.

Subbaramaiah K, Simms S . Photolabeling of CheR methyltransferase with S-adenosyl-L-methionine (AdoMet). Studies on the AdoMet binding site. J Biol Chem. 1992; 267(12):8636-42. View

16.

Roessner C, Warren M, Santander P, Atshaves B, Ozaki S, Stolowich N . Expression of 9 Salmonella typhimurium enzymes for cobinamide synthesis. Identification of the 11-methyl and 20-methyl transferases of corrin biosynthesis. FEBS Lett. 1992; 301(1):73-8. DOI: 10.1016/0014-5793(92)80213-z. View

17.

Goldman B, Roth J . Genetic structure and regulation of the cysG gene in Salmonella typhimurium. J Bacteriol. 1993; 175(5):1457-66. PMC: 193233. DOI: 10.1128/jb.175.5.1457-1466.1993. View

18.

Roth J, Lawrence J, Rubenfield M, Church G . Characterization of the cobalamin (vitamin B12) biosynthetic genes of Salmonella typhimurium. J Bacteriol. 1993; 175(11):3303-16. PMC: 204727. DOI: 10.1128/jb.175.11.3303-3316.1993. View

19.

Debussche L, THIBAUT D, Cameron B, Crouzet J, Blanche F . Biosynthesis of the corrin macrocycle of coenzyme B12 in Pseudomonas denitrificans. J Bacteriol. 1993; 175(22):7430-40. PMC: 206888. DOI: 10.1128/jb.175.22.7430-7440.1993. View

20.

Spencer J, Stolowich N, Roessner C, Scott A . The Escherichia coli cysG gene encodes the multifunctional protein, siroheme synthase. FEBS Lett. 1993; 335(1):57-60. DOI: 10.1016/0014-5793(93)80438-z. View