The pterin component of the molybdenum factor. Structural characterization of two fluorescent derivatives

J. L. Johnson, Bryan Hainline, K. V. Rajagopalan, B. H. Arison

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Abstract

Two stable fluorescent derivatives of molybdopterin have been structurally characterized. Form A is an oxidized pterin with a 6-alkyl substituent. Results of chemical, mass spectral, and NMR studies are consistent with the side chain formulation -C≡C-CH-OHCH2OPO32-. Similar studies on the Form B derivative indicate that it is the phosphorylated analog of urothione but lacks the 3-methylthio function. Form B (dephospho) can be synthesized from urothione by desulfuration with Raney nickel and oxidation with SeO2. Chicken liver sulfite oxidase (sulfite:ferricytochrome c oxidoreductase, EC 1.8.2.1) contains one phosphate residue (as molybdopterin) per subunit. The phosphate is noncovalently bound but is not released by trichloroacetic acid at 4°C. The yield of Form A and Form B from sulfite oxidase is 0.50 and 0.27/subunit, respectively. The phosphate ester bond in isolated molybdopterin (Form B) is partially hydrolyzed by 1 N HCl at 100°C (33% in 1 h). The release of inorganic phosphate from sulfite oxidase was more rapid (35% in 10 min) due to the presence of molybdate in the denatured enzyme mixture but slower than expected from a high energy phosphate bond. The presence of molybdopterin in a wide variety of molybdenum- and tungsten-containing enzymes has been demonstrated. Glucose oxidase and the iron and manganese superoxide dismutases are devoid of molybdopterin.

Original languageEnglish (US)
Pages (from-to)5414-5422
Number of pages9
JournalJournal of Biological Chemistry
Volume259
Issue number9
StatePublished - 1984
Externally publishedYes

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Pterins
Sulfites
Sulfite Dehydrogenase
Molybdenum
Derivatives
Cytochromes c
Phosphates
Oxidoreductases
Superoxide Dismutase
Trichloroacetic Acid
Tungsten
Enzymes
Nickel
Liver
Chickens
Esters
Nuclear magnetic resonance
molybdenum cofactor
Glucose
Oxidation

ASJC Scopus subject areas

  • Biochemistry

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The pterin component of the molybdenum factor. Structural characterization of two fluorescent derivatives. / Johnson, J. L.; Hainline, Bryan; Rajagopalan, K. V.; Arison, B. H.

In: Journal of Biological Chemistry, Vol. 259, No. 9, 1984, p. 5414-5422.

Research output: Contribution to journalArticle

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