Gln5 selectively monodansylated substance P as a sensitive tool for interaction studies with membranes.
Substance P (SP) is a neuropeptide endowed with several important biological activities both in the central and peripheral nervous system. Taking advantage of the presence of glutamine residues in SP, the peptide was labelled with the fluorescent probe monodansylcadaverine using the transglutaminase (TGase)-reaction in order to study interactions between SP and model or natural membranes. Although it was verified that both adjacent glutamines of the peptide can act as substrate for TGase in a consecutive reaction, conditions were optimized to selectively label Gln5. This fluorescent SP analogue was found to adopt environment-dependent conformations similar to those of the natural peptide and proved to be functionally active on guinea pig trachea. Fluorescence spectroscopy was used to demonstrate the potential use of dansylated SP in studies involving interactions with membranes.
Ferrándiz
C
C
Departament de Bioquímica i Biologia Molecular, Universitat de València, Spain.
Pérez-Payá
E
E
Braco
L
L
Abad
C
C
D000595:Amino Acid Sequence; D000818:Animals; D002103:Cadaverine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D005973:Glutamine; D006168:Guinea Pigs; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D010743:Phospholipids; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013373:Substance P / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000378:metabolism; D011503:Transglutaminases / Q000378:metabolism*
D010743:Phospholipids; D005973:Glutamine; D013373:Substance P; D011503:Transglutaminases; D002103:Cadaverine