title: Solid-phase synthesis and purification of a set of uniformly 13C, 15N labelled de novo designed membrane fusogenic peptides.
abstract: The transmembrane segments of soluble N-ethylmaleimide-sensitive factor (SNARE) proteins or viral envelope proteins drive membrane fusion, which suggests that simple synthetic biology constructs for fusion exist and can be evaluated. We describe the high-yield synthesis of a set of de novo designed fusogenic peptides for use in functional investigations, which are highly enriched in 13C and 15N using three equivalents of labelled amino acids and optimized reaction conditions minimizing aggregation. The biomimetic peptides have a high purity >90% and show reproducible and fusogenic activity that correlates well with the intended functional design characteristics, from strongly fusogenic to almost non-fusogenic.
authors.affiliation: Biophysical Organic Chemistry, Solid State NMR, Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands.
authors.forename: Prashant R
authors.initials: PR
authors.lastname: Agrawal
authors.forename: Mathias W
authors.initials: MW
authors.lastname: Hofmann
authors.forename: Nico J
authors.initials: NJ
authors.lastname: Meeuwenoord
authors.forename: Dmitri V
authors.initials: DV
authors.lastname: Filippov
authors.forename: Holger
authors.initials: H
authors.lastname: Stalz
authors.forename: Frans
authors.initials: F
authors.lastname: Hulsbergen
authors.forename: Dieter
authors.initials: D
authors.lastname: Langosch
authors.forename: Hermen S
authors.initials: HS
authors.lastname: Overkleeft
authors.forename: Huub
authors.initials: H
authors.lastname: De Groot
mesh_terms: D000595:Amino Acid Sequence; D002247:Carbon Isotopes; D005089:Exocytosis; D007553:Isotope Labeling / Q000379:methods*; D008081:Liposomes; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000138:chemical synthesis / Q000502:physiology; D009587:Nitrogen Isotopes; D010455:Peptides / Q000138:chemical synthesis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D009435:Synaptic Transmission
chemical_list: D002247:Carbon Isotopes; D008081:Liposomes; D009419:Nerve Tissue Proteins; D009587:Nitrogen Isotopes; D010455:Peptides