Leveraging orthogonal mass spectrometry based strategies for comprehensive sequencing and characterization of ribosomal antimicrobial peptide natural products

Abstract

Strategies to accelerate natural product peptide characterization.

Post‐Translational Formation of Aminomalonate by a Promiscuous Peptide‐Modifying Radical SAM Enzyme

“…[31,38] 1, SjiA 51-60 -2 exhibited as eries of ba nd y ions within the tobe-cyclized FxS motif (Figure 2E and Figure S3), indicating that the FxS motif was not cross-linked in 2.M ore importantly,t he À2Dam odification was observed exclusively on the Ser residue within the FxS motif (Figure 2E), which is reminiscent of formylglycine (FGly), astructural motif can be formed by anaerobic sulfatase maturation enzyme (anSME). [40][41][42] This proposal is consistent with the observation of the + 16 Da Angewandte Chemie modifications, [43] but is reminiscent of Ama, am alonatecontaining amino acid. [44,45] We hence speculated that SjiA 51-60 -4 contains aSer-derived Ama, which was decarboxylated to produce aGly during collision-induced dissociation (CID) in HR-MS/MS analysis.C onsistent with this proposal, the HR-MS/MS spectrum of SjiA 51-60 -4 matches well to aS er-to-Gly mutant of SjiA 51-60 (i.e.t he ion with [M+ +H] + = 1006.52), in which the Ser residue in the FxS motif is changed to aG ly.…”

“…Chemie modifications, [43] but is reminiscent of Ama, am alonatecontaining amino acid. [44,45] We hence speculated that SjiA 51-60 -4 contains aSer-derived Ama, which was decarboxylated to produce aGly during collision-induced dissociation (CID) in HR-MS/MS analysis.C onsistent with this proposal, the HR-MS/MS spectrum of SjiA 51-60 -4 matches well to aS er-to-Gly mutant of SjiA 51-60 (i.e.t he ion with [M+ +H] + = 1006.52), which the Ser residue in the FxS motif is changed to aG ly.…”

Post‐Translational Formation of Aminomalonate by a Promiscuous Peptide‐Modifying Radical SAM Enzyme

“…m / z= 1050.52), suggesting a decarboxylation‐type neutral loss in MS/MS analysis (Figure 2 F and Figure S5). This type of neutral loss has not been observed for peptidyl moiety produced by common post‐translational modifications, [43] but is reminiscent of Ama, a malonate‐containing amino acid [44, 45] . We hence speculated that SjiA 51 – 60 ‐4 contains a Ser‐derived Ama, which was decarboxylated to produce a Gly during collision‐induced dissociation (CID) in HR‐MS/MS analysis.…”

“…Chemie Forschungsartikel 20112 www.angewandte.de modifications, [43] but is reminiscent of Ama, am alonatecontaining amino acid. [44,45] We hence speculated that SjiA 51-60 -4 contains aSer-derived Ama, which was decarboxylated to produce aGly during collision-induced dissociation (CID) in HR-MS/MS analysis.C onsistent with this proposal, the HR-MS/MS spectrum of SjiA 51-60 -4 matches well to aS er-to-Gly mutant of SjiA 51-60 (i.e.t he ion with [M+ +H] + = 1006.52), in which the Ser residue in the FxS motif is changed to aG ly.…”

Tuning the Activity of Anoplin by Dendrimerization of Lysine and Lipidation of the N-Terminal

“…In conclusion, in the development of AMPs, excessive chemical modifications may not be necessary for some peptides. In contrast, simple modifications may lead to unexpected and satisfactory results. − The direct lipidation of a single anoplin is better than the lipidation after dendrimerization for antibacterial performance and cell selectivity. Dendrimerization increases the hydrophobicity, cationic nature, α-helicity, and multi-valency of these compounds, thereby increasing the antibacterial activity.…”