DOI: 10.1128/jb.169.1.110-116.1987. In vitro system for molybdopterin biosynthesis

In vitro system for molybdopterin biosynthesis

10.1128/jb.169.1.110-116.1987

Crossref journal-article

American Society for Microbiology

Journal of Bacteriology (235)

Abstract

A high-Mr fraction present in chl+ and chlA1 strains of Escherichia coli synthesizes molybdopterin (MPT) from the low-Mr fraction of several MPT-deficient mutants. Using this in vitro complementation as an assay, we have partially characterized the high-Mr fraction as a protein, termed MPT converting factor, of Mr 45,000, distinguishable from the Mo cofactor carrier protein of similar Mr by its absolute requirement for the low-Mr fraction of a non-chlA1 mutant in the nit-1 reconstitution assay. MPT converting factor was rapidly inactivated in the absence of a reduced sulfhydryl compound. Anaerobic incubation of MPT converting factor with trypsin destroyed its activity. High-performance liquid chromatographic analysis of alkaline KMnO4 oxidation products demonstrated that the factor did not contain any bound pterin. Since mutants lacking MPT converting factor are not auxotrophs for folate or riboflavin, the factor appears to be distinct from known pteridine biosynthetic enzymes in E. coli. We have partially purified and characterized the low-Mr fractions as probable MPT precursors. Several distinct precursors were separable by high-performance liquid chromatography. Like MPT activity, precursor activity was oxygen sensitive. Precursor activity was not correlated with levels of L-threo-neopterin, a major pterin of unknown function in E. coli. Precursor activity was correlated with levels of a new 6-alkylpterin, compound Z, produced by acidic iodine oxidation. Compound Z has the properties expected of an oxidized MPT precursor.

Bibliography

Johnson, M. E., & Rajagopalan, K. V. (1987). In vitro system for molybdopterin biosynthesis. Journal of Bacteriology, 169(1), 110â116.

Authors 2

M E Johnson (first)
K V Rajagopalan (additional)

References 15 Referenced 38

Amy , N. K. 1981 . Identification of the molybdenum cofactor in chlorate-resistant mutants of Escherichia coli . J. Bacteriol. 148 : 274 - 282 . (10.1128/jb.148.1.274-282.1981) / J. Bacteriol. / Identification of the molybdenum cofactor in chlorate-resistant mutants of Escherichia coli by Amy N. K. (1981)
Amy , N. K. , and K. V. Rajagopalan . 1979 . Characterization of molybdenum cofactor from Escherichia coli . J. Bacteriol. 140 : 114 - 124 . (10.1128/jb.140.1.114-124.1979) / J. Bacteriol. / Characterization of molybdenum cofactor from Escherichia coli by Amy N. K. (1979)
Blakley R. L. 1969. The biochemistry of folic acid and related pteridines. p. 58-105 North-Holland Publishing Co. Amsterdam. %
Brown , G. M. , and J. M. Williamson . 1982 . Biosynthesis of riboflavin, folic acid, thiamine, and pantothenic acid . Adv. Enzymol. 53 : 345 - 381 . / Adv. Enzymol. / Biosynthesis of riboflavin, folic acid, thiamine, and pantothenic acid by Brown G. M. (1982)
Fukushima , T. , and J. C. Nixon . 1980 . Analysis of reduced forms of biopterin in biological tissues and fluids . Anal. Biochem. 102 : 176 - 188 . (10.1016/0003-2697(80)90336-X) / Anal. Biochem. / Analysis of reduced forms of biopterin in biological tissues and fluids by Fukushima T. (1980)
Giordano , G. , L. Saracino , and L. Grillet . 1985 . Identification in various chlorate-resistant mutants of a protein involved in the activation of nitrate reductase in the soluble fraction of a chlA mutant of Escherichia coli K-12 . Biochim. Biophys. Acta 839 : 181 - 190 . (10.1016/0304-4165(85)90035-2) / Biochim. Biophys. Acta / Identification in various chlorate-resistant mutants of a protein involved in the activation of nitrate reductase in the soluble fraction of a chlA mutant of Escherichia coli K-12 by Giordano G. (1985)
Grillet , L. , and G. Giordano . 1983 . Identification and purification of a protein involved in the activation of nitrate reductase in the soluble fraction of a chiA mutant of Escherichia coli K12 . Biochim. Biophys. Acta 749 : 115 - 124 . (10.1016/0167-4838(83)90158-9) / Biochim. Biophys. Acta / Identification and purification of a protein involved in the activation of nitrate reductase in the soluble fraction of a chiA mutant of Escherichia coli K12 by Grillet L. (1983)
Hefti , F. 1982 . High-performance size-exclusion chromatography: a buffer for the reliable determination of molecular weights of proteins . Anal. Biochem. 121 : 378 - 381 . (10.1016/0003-2697(82)90496-1) / Anal. Biochem. / High-performance size-exclusion chromatography: a buffer for the reliable determination of molecular weights of proteins by Hefti F. (1982)
Heine , M. C. , and G. M. Brown . 1975 . Enzymatic epimerization of D-erythro-dihydroneopterin triphosphate to L-threodihydroneopterin triphosphate . Biochim. Biophys. Acta 411 : 236 - 249 . (10.1016/0304-4165(75)90304-9) / Biochim. Biophys. Acta / Enzymatic epimerization of D-erythro-dihydroneopterin triphosphate to L-threodihydroneopterin triphosphate by Heine M. C. (1975)
Johnson , J. L. , and K. V. Rajagopalan . 1982 . Structural and metabolic relationship between the molybdenum cofactor and urothione . Proc. Natl. Acad. Sci. USA 79 : 6856 - 6860 . (10.1073/pnas.79.22.6856) / Proc. Natl. Acad. Sci. USA / Structural and metabolic relationship between the molybdenum cofactor and urothione by Johnson J. L. (1982)
Johnson , J. L. , B. E. Hainline , K. V. Rajagopalan , and B. H. Arison . 1984 . The pterin component of the molybdenum cofactor: structural characterization of two fluorescent derivatives . J. Biol. Chem. 259 : 5414 - 4522 . (10.1016/S0021-9258(18)91027-6) / J. Biol. Chem. / The pterin component of the molybdenum cofactor: structural characterization of two fluorescent derivatives by Johnson J. L. (1984)
Johnson , M. E. , and K. V. Rajagopalan . 1986 . Involvement of chlA, E, M, and N loci in Escherichia coli molybdopterin biosynthesis . J. Bacteriol. 169 : 117 - 125 . (10.1128/jb.169.1.117-125.1987) / J. Bacteriol. / Involvement of chlA, E, M, and N loci in Escherichia coli molybdopterin biosynthesis by Johnson M. E. (1986)
Kramer , S. , R. V. Hageman , and K. V. Rajagopalan . 1984 . In vitro reconstitution of nitrate reductase activity of the Neurospora crassa mutant nit-i: specific incorporation of molybdopterin . Arch. Biochem. Biophys. 233 : 821 - 829 . (10.1016/0003-9861(84)90511-3) / Arch. Biochem. Biophys. / In vitro reconstitution of nitrate reductase activity of the Neurospora crassa mutant nit-i: specific incorporation of molybdopterin by Kramer S. (1984)
Rembold , H. , and G. Heinz . 1971 . L-threo-neopterin, the major pterin in Escherichia coli B . Z. Phys. Chem. 352 : 1271 - 1272 . / Z. Phys. Chem. / L-threo-neopterin, the major pterin in Escherichia coli B by Rembold H. (1971)
Richey , D. P. , and G. M. Brown . 1969 . The biosynthesis of folic acid. IX. Purification and properties of the enzymes required for the formation of dihydropteroic acid . J. Biol. Chem. 244 : 1582 - 1592 . / J. Biol. Chem. / The biosynthesis of folic acid. IX. Purification and properties of the enzymes required for the formation of dihydropteroic acid by Richey D. P. (1969)

Dates

Type	When
Created	8 years, 10 months ago (Nov. 2, 2016, 4:28 p.m.)
Deposited	4 years, 1 month ago (July 29, 2021, 1:34 p.m.)
Indexed	1 year, 10 months ago (Oct. 25, 2023, 5:59 p.m.)
Issued	38 years, 8 months ago (Jan. 1, 1987)
Published	38 years, 8 months ago (Jan. 1, 1987)
Published Print	38 years, 8 months ago (Jan. 1, 1987)

Funders 0

None

BibTeX

@article{Johnson_1987, title={In vitro system for molybdopterin biosynthesis}, volume={169}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.169.1.110-116.1987}, DOI={10.1128/jb.169.1.110-116.1987}, number={1}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Johnson, M E and Rajagopalan, K V}, year={1987}, month=jan, pages={110–116} }

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