252Cf-plasma desorption mass spectrometry analysis of lipids A obtained by an elimination reaction under mild conditions.

Lipids A are the hydrophobic domains of bacterial endotoxic lipopolysaccharides. Since they are responsible for most of the biological activities (both pathogenic and beneficial) of endotoxins, the characterization of their structure is crucial to the understanding of their mode of action. However, the inadequacy of existing methods for preparing certain lipids A has prompted us to devise a new, mild procedure which gives intact products. Use was made of the special features of 252Cf-plasma desorption mass spectrometry for forming molecular ions from these species and giving qualitative and quantitative information from the primary mass spectrum.

Plasma desorption mass spectrometry of two synthetic sarafotoxins: side reactions and characterization of the intermediates.

Sarafotoxins (SRTXs) form a family of toxic and potent vasoconstrictor peptides of 21 amino acids and two disulfide bonds. They are present in the venom of the burrowing asp Atractaspis engaddensis. We have made two derivatives of the amino acid sequence of SRTX-b, one of the most potent isotoxins, in the solid phase. First, we replaced Ser2 by Thr, to investigate whether, as previously postulated, this change is responsible for the weak activities of SRTXs c and d. Secondly, we replaced Ser2, Asp18 and Val19 respectively by Thr, Gly and Ile, with a view to generating SRTX-e whose amino acid sequence was deduced from cDNA. Solid-phase peptide synthesis (SPPS) was performed according to the tert-butyloxycarbonyl strategy and the disulfides were paired sequentially using a selective chemistry. The disulfide 1-15 was formed by oxidation of cysteines1,15 with ferricyanide, whereas disulfide 3-11 was made by iodine oxidation of Acm-blocked cysteines3,11. By plasma desorption mass spectrometry (PDMS), we monitored all possible side reactions that occurred during the synthesis. We thus observed a benzyl shift in mass spectra when aspartic and glutamic acid side chains were protected by a benzyl group during the SPPS. This could be circumvented by using instead, a cyclohexyl protecting group. We also noted the oxidation of the methionine and the tryptophan side chain (formation of methionine sulfoxide and oxindole ring of tryptophan) to a small extent during the cleavage peptide/solid phase oxidation of the methionine side chain during the formation of the disulfide 1-15 by ferricyanide.(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of sequence on the fragmentation of serine- and threonine-containing peptides in 252Cf-plasma desorption mass spectrometry.

The molecular weights of four linear synthetic peptides, fragments of a snake alpha-neurotoxin, were measured by 252Cf-plasma desorption mass spectrometry. The fragmentation phenomenon observed at the level of serine and/or threonine residue with a concomitant ion/fragment association is reported for a group of two peptides (B and D) in contrast with the group (A and C) in spite of the high ratio of serine and threonine, namely peptide A. The propensity for specific fragmentation of peptide D seems to be correlated to the repetitive sequence, (Gly-Ser)2. Finally, based on the m/z of the daughter-ions measured, we propose an overall mechanism as an N----O acyl shift analogous to that observed for serine- and threonine- containing peptides in solution chemistry.

Identification of dekamycin antibiotics.

Dekamycin was isolated from strain DK5 of Streptomyces fradiae, and purified by ion-exchange column chromatography. Excellent separation of N-acetylated derivatives of the antibiotic mixture was achieved by two different methods, flash chromatography and high-performance liquid chromatography. Mass spectrometry (252Cf plasma desorption mass spectrometry, fast atom bombardment) for the analysis of N-acetyldekamycins (neomycin B, neomycin C, neamine and ribostamycin) was mainly used to confirm the molecular weight of a structure deduced by 1H and 13C NMR analyses.