Synthesis, lipophilic derivatization and interaction with liposomes of HAV-VP3 (102-121) sequence by using spectroscopic techniques.
Hepatitis A virus (HAV) is composed mainly of three structural capsid proteins: VP1, VP2 and VP3. Our group has reported the synthesis and the immunogenic evaluation of VP3 (110-121) peptide sequence. In the present work, in order to stimulate a T-cell immune response, we have selected the HAV-VP3 (102-121) peptide which has maximum amphipathicity. Its synthesis was carried out manually in the solid phase and semipreparative HPLC was used for purification of the crude peptide. Finally the purified peptide was characterized by analytical HPLC, amino acid analysis and MS. A palmitoyl derivative of VP3 (102-121) was synthesized to modify the hydrophobicity of the peptide. Both free and lipophilically derivatized peptides were incorporated into multilamelar liposomes. Physicochemical studies of the HAV-related peptides described above were carried out using monolayers as membrane models. Compression isotherms, surface activity and penetration kinetics into dipalmitoylphosphatidylcholine monolayers were determined. Moreover, changes in the fluidity of bilayers induced by these peptides were determined by means of polarizable probes such as 8-anilino-1-naphthalenesulfonic acid and 1,6-diphenyl-1,3,5-hexatriene. The integrity of the membranes has also been ascertained with the carboxyfluorescein.
Garcia
M
M
Department of Peptide and Protein Chemistry, CID, CSIC, Barcelona, Spain.
Pujol
M
M
Reig
F
F
Alsina
M A
MA
Haro
I
I
D000595:Amino Acid Sequence; D002213:Capsid / Q000737:chemistry; D005454:Fluorescence Polarization; D006507:Hepatovirus / Q000737:chemistry*; D008081:Liposomes; D008560:Membrane Fluidity; D008969:Molecular Sequence Data; D012995:Solubility; D013816:Thermodynamics; D014764:Viral Proteins / Q000737:chemistry*
D008081:Liposomes; D014764:Viral Proteins