Aggregation, fusion and aqueous content release from liposomes induced by lysozyme derivatives: effect on the lytic activity.
Chemically modified lysozymes, namely: N-succinyl lysozyme, glycine methyl ester of N-succinyl lysozyme and oxoindole lysozyme have been prepared. Aggregation, fusion and leakage of phospholipid vesicles induced by these derivatives have been studied in comparison with the effect of the unmodified protein. The experiments were carried out with negatively charges 9PC/PA, 9:1) and uncharged (PC and PC/DOPE/Chol (10:5:5)) lipid vesicles of different packing. Fusion and aggregation of negatively charged phospholipid vesicles in induced by proteins positively charged at pH 7.0 involving electrostatic interactions, a similar pattern on fusion and aggregation of the least stably packed lipid vesicles points also to hydrophobic forces playing a role in the lipid-protein interaction. A conformational change of the protein involved increasing beta-turns, loops and unordered structure at the expenses of beta-sheet without affecting alpha helix content. The conformational effect is necessary to provoke the effects studied, since one of the derivatives (N-succinyl lysozyme) neither changes conformation nor causes aggregation and fusion of vesicles. However, there is no relationship between lysozyme activity and fusion or aggregation of lipid vesicles that catalytic and fusogenci sites of, indicating lysozyme are topographically different.
Vechetti
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GF
Departamento Bioquímica de la Nutrición, Instituto de Química Biológica Dr. B. Bloj, Facultad de Bioquímica, Química y Farmacia, (CONICET-UNT) 4000 San Miguel de Tucamán, Argentina.
de Arcuri
B F
BF
Posse
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Arrondo
J L
JL
Morero
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RD
D000818:Animals; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002784:Cholesterol / Q000737:chemistry; D005998:Glycine / Q000031:analogs & derivatives / Q000494:pharmacology; D008081:Liposomes / Q000378:metabolism*; D008561:Membrane Fusion / Q000187:drug effects; D009113:Muramidase / Q000737:chemistry / Q000494:pharmacology*; D010539:Permeability / Q000187:drug effects; D010712:Phosphatidic Acids / Q000737:chemistry; D010713:Phosphatidylcholines / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry; D011487:Protein Conformation; D013386:Succinates / Q000494:pharmacology
D008081:Liposomes; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D013386:Succinates; C094877:dioleoyl phosphatidylethanolamine; D002784:Cholesterol; C110021:N-succinyl lysozyme; C110019:glycine methyl ester lysozyme; C110016:oxoindole lysozyme; D009113:Muramidase; D005998:Glycine