The structure of U17 isolated from Streptomyces clavuligerus and its properties as an antioxidant thiol.
The predominant low-molecular-mass thiol produced by streptomycetes is a cysteine derivative previously designated as U17 [Newton, G. L., Fahey, R. C., Cohen, G. & Aharonowitz, Y. (1993) J. Bacteriol. 175, 2734-2742]. In this study we report the elucidation of the structure of the monobromobimane derivative of U17, which establishes the structure of U17 as 2-(N-acetylcysteinyl)amido-2-deoxy-alpha-D-glucopyranosyl-myo-inositol. The presence of the N-acetylcysteine moiety was indicated by formation of N-acetylcysteine-monobromobimane during acid hydrolysis of the monobromobimane derivative of U17. Complete hydrolysis released 1 mol glucosamine/mol cysteine as determined by carbohydrate and amino acid analysis. High-resolution mass spectral analysis gave a precise mass consistent with the molecular formula C27H40N4O14S. Analysis of 13C-NMR, one-dimensional 1H-NMR and two-dimensional NMR experiments identified the remaining C6H12O6 moiety as myo-inositol, confirmed the presence of N-acetylcysteine and glucosamine, and established the connectivity of the components. Two chemical properties of this novel thiol make it suitable as an intracellular storage form of cysteine and as an antioxidant thiol. First, it undergoes heavy-metal-ion catalyzed autoxidation at a rate dramatically lower than that for cysteine and markedly lower than that for glutathione or N-acetylcysteine. Secondly, the alpha-(1-->1) glycosidic link between glucosamine and myo-inositol is resistant to acid hydrolysis, hydrolysing at a rate comparable to that of the two amide bonds in the molecule.
Newton
G L
GL
Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla 92093-0506, USA.
Bewley
C A
CA
Dwyer
T J
TJ
Horn
R
R
Aharonowitz
Y
Y
Cohen
G
G
Davies
J
J
Faulkner
D J
DJ
Fahey
R C
RC
D000975:Antioxidants / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine; D004187:Disaccharides / Q000737:chemistry* / Q000494:pharmacology; D006020:Glycopeptides; D007294:Inositol; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011720:Pyrazoles; D013302:Streptomyces / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000737:chemistry* / Q000494:pharmacology
D000975:Antioxidants; D004187:Disaccharides; D006020:Glycopeptides; D011720:Pyrazoles; D013438:Sulfhydryl Compounds; C089265:mycothiol; D007294:Inositol; D003545:Cysteine