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. 1986 Feb;83(4):962–966. doi: 10.1073/pnas.83.4.962

Lysozyme-induced fusion of liposomes with erythrocyte ghosts at acidic pH.

T Arvinte, K Hildenbrand, P Wahl, C Nicolau
PMCID: PMC322990  PMID: 3456575

Abstract

Lysozyme that was covalently bound to the outer surface of sonicated vesicles induced fusion of the vesicles with human white erythrocyte ghosts. The kinetics of membrane mixing were evaluated by the resonance-energy-transfer method using L-alpha-dipalmitoyl phosphatidylethanolamine labeled at the free amino group with the energy donor 7-nitro-2,1,3-benzoxadiazol-4-yl or with the energy acceptor tetramethylrhodamine. The equilibrium state after fusion was characterized by using fluorescence photobleaching and recovery techniques. Rates and equilibrium percentages of fusion were maximal at the pH optimum of the enzyme, and rates were strongly reduced by the addition of N,N',N''-triacetylchitotriose, a competitive inhibitor of lysozyme. An apparent activation energy of 28 +/- kcal/mol was obtained for the lipid-mixing process. At 37 degrees C, the fusion half-time was 0.5 min. After 30 min at 37 degrees C, 40% of the labeled lipids initially present in the fusion mixture had a lateral diffusion constant, D, of 1.1 +/- 0.5 X 10(-9) cm2 X sec-1 in the ghost membrane. The strong induction of fusion at the lysozyme pH optimum was not observed in the absence of lysozyme or when free lysozyme was added to the solution. Bound lysozyme did not induce fusion of electrically neutral liposomes with each other. These observations indicate that it is the liposome-bound lysozyme that induces fusion between liposomes and erythrocyte ghosts.

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Selected References

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