Isolation, mass spectrometric characterization, and protein phosphatase inhibition properties of cyclic peptide analogues of gramicidin-S from Bacillus brevis (Nagano strain).
Six substitutional variants of gramicidin-S, present in a commercial extract of Bacillus brevis, have been characterized using a combination of liquid chromatographic/mass spectrometric analysis of the crude extract, plus accurate mass measurements, tandem mass spectrometric investigations, and amino acid analyses, of isolated high-performance liquid chromatographic fractions. Two of these variants were shown previously (Nozaki and Muramatsu, J. Antibiotics 37, 689 (1984)) to involve replacement of one or both Val residues in gramicidin-S by aminobutyric acid, and these findings are confirmed here. One of the four unknown variants corresponds to substitution of one Val residue by Leu, while the other three all involve substitution of one of the Orn residues in gramicidin-S by Lys, or by citrulline (gamma-carbamoyl ornithine), or by gamma-formyl ornithine. The biological activities of the purified fractions were assessed with respect to their in vitro inhibition of protein phosphatases PP1 and PP2A. For both protein phosphatases inhibition levels fell in the micromolar range, about four orders of magnitude higher than IC50 values for other small cyclic peptides such as microcystins and nodularins.
Thibault
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P
Institute for Marine Biosciences, National Research Council, Halifax, Nova Scotia, Canada.
Faubert
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D
Karunanithy
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S
Boyd
R K
RK
Holmes
C F
CF
D000596:Amino Acids / Q000032:analysis; D001407:Bacillus / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D006096:Gramicidin / Q000737:chemistry* / Q000494:pharmacology; D013058:Mass Spectrometry; D010455:Peptides / Q000302:isolation & purification* / Q000494:pharmacology; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors*; D013056:Spectrophotometry, Ultraviolet
D000596:Amino Acids; D010455:Peptides; D006096:Gramicidin; D010744:Phosphoric Monoester Hydrolases