New high-performance liquid chromatography-based methodology for monitoring the conformational transitions of self-associating hydrophobic peptides, incorporated into liposomes.
A new high-performance size-exclusion chromatographic strategy is reported for the analysis of the hydrophobic self-associating peptide gramicidin A, incorporated into artificial phospholipid vesicles (liposomes). The method is based on the direct injection of a few microlitres of the gramicidin A-containing liposome suspension into the column, which is eluted with a non-polar solvent, such as tetrahydrofuran. The type and amount of information which can be derived from this methodology have been evaluated. Using this chromatographic approach, a correlation has been unambiguously shown to exist between the organization of the peptide in the vesicles and a number of variables involved in the method of preparation of liposomes. Finally, a gramicidin A conformational transition has been monitored in the phospholipid vesicles which proved to be dependent on the class of phospholipid present in the liposome.
Bañó
M C
MC
Departament de Bioquímica i Biología Molecular, Facultades de Ciencias, Universitat de Valencia, Spain.
Braco
L
L
Abad
C
C
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006096:Gramicidin / Q000032:analysis; D007700:Kinetics; D008081:Liposomes / Q000032:analysis*; D010455:Peptides / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet
D008081:Liposomes; D010455:Peptides; D006096:Gramicidin