Solid-phase synthesis and purification of a set of uniformly 13C, 15N labelled de novo designed membrane fusogenic peptides.
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.
Agrawal
Prashant R
PR
Biophysical Organic Chemistry, Solid State NMR, Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands.
Hofmann
Mathias W
MW
Meeuwenoord
Nico J
NJ
Filippov
Dmitri V
DV
Stalz
Holger
H
Hulsbergen
Frans
F
Langosch
Dieter
D
Overkleeft
Hermen S
HS
De Groot
Huub
H
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
D002247:Carbon Isotopes; D008081:Liposomes; D009419:Nerve Tissue Proteins; D009587:Nitrogen Isotopes; D010455:Peptides