Signatures of Mechanically Interlocked Topology of Lasso Peptides by Ion Mobility-Mass Spectrometry: Lessons from a Collection of Representatives.
Lasso peptides are characterized by a mechanically interlocked structure, where the C-terminal tail of the peptide is threaded and trapped within an N-terminal macrolactam ring. Their compact and stable structures have a significant impact on their biological and physical properties and make them highly interesting for drug development. Ion mobility - mass spectrometry (IM-MS) has shown to be effective to discriminate the lasso topology from their corresponding branched-cyclic topoisomers in which the C-terminal tail is unthreaded. In fact, previous comparison of the IM-MS data of the two topologies has yielded three trends that allow differentiation of the lasso fold from the branched-cyclic structure: (1) the low abundance of highly charged ions, (2) the low change in collision cross sections (CCS) with increasing charge state and (3) a narrow ion mobility peak width. In this study, a three-dimensional plot was generated using three indicators based on these three trends: (1) mean charge divided by mass (ζ), (2) relative range of CCS covered by all protonated molecules (ΔΩ/Ω) and (3) mean ion mobility peak width (δΩ). The data were first collected on a set of twenty one lasso peptides and eight branched-cyclic peptides. The indicators were obtained also for eight variants of the well-known lasso peptide MccJ25 obtained by site-directed mutagenesis and further extended to five linear peptides, two macrocyclic peptides and one disulfide constrained peptide. In all cases, a clear clustering was observed between constrained and unconstrained structures, thus providing a new strategy to discriminate mechanically interlocked topologies. Graphical Abstract ᅟ.
Normandie Univ, UNIROUEN, INSA Rouen, CNRS, COBRA, 76000, Rouen, France.
Kevin Jeanne Dit
KJD
Fouque
Normandie Univ, UNIROUEN, INSA Rouen, CNRS, COBRA, 76000, Rouen, France. helene.lavanant@univ-rouen.fr.
Hélène
0000-0002-2963-0327
H
Lavanant
Muséum National d'Histoire Naturelle, Sorbonne Universités, Centre national de la Recherche scientifique, Laboratoire Molécules de Communication et Adaptation des Microorganismes, UMR 7245 CNRS-MNHN, CP 54, 57 rue Cuvier, 75005, Paris, France.
Séverine
S
Zirah
Department of Chemistry, Biochemistry; LOEWE Center for Synthetic Microbiology, Philipps-University Marburg, Hans-Meerwein-Strasse 4, 35032, Marburg, Germany.
Julian D
JD
Hegemann
Department of Chemistry, Biochemistry; LOEWE Center for Synthetic Microbiology, Philipps-University Marburg, Hans-Meerwein-Strasse 4, 35032, Marburg, Germany.
Marcel
M
Zimmermann
Department of Chemistry, Biochemistry; LOEWE Center for Synthetic Microbiology, Philipps-University Marburg, Hans-Meerwein-Strasse 4, 35032, Marburg, Germany.
Mohamed A
MA
Marahiel
Muséum National d'Histoire Naturelle, Sorbonne Universités, Centre national de la Recherche scientifique, Laboratoire Molécules de Communication et Adaptation des Microorganismes, UMR 7245 CNRS-MNHN, CP 54, 57 rue Cuvier, 75005, Paris, France.
Sylvie
S
Rebuffat
Normandie Univ, UNIROUEN, INSA Rouen, CNRS, COBRA, 76000, Rouen, France.
Carlos
C
Afonso
D001430:Bacteriocins / Q000737:chemistry / Q000235:genetics; D004220:Disulfides / Q000737:chemistry; D000075663:Ion Mobility Spectrometry / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010455:Peptides / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation
D001430:Bacteriocins; D004220:Disulfides; D010455:Peptides; C533129:capistruin; C020411:microcin
Charge state distribution; Collision cross section range; Constrained peptides; Disulfide bond; Ion mobility; Ion mobility peak width; Lasso peptides; Macrocyclic peptide; Mechanically interlocked peptides