The lipid integrity of membranes of guinea pig alveolar macrophages studied by nanosecond fluorescence decay of 1,6-diphenyl-1,3,5-hexatriene: The effect of stimulation by concanavalin A and formyl peptides1

S. N. Cherenkevich, J. M. Vanderkooi, R. Restifo, R. P. Daniele, A. Holian

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Abstract

Time-resolved fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene was used to monitor physical changes in the membranes of guinea pig alveolar macrophages following stimulation by N-formyl peptides (either N-formylmethionylphenylalanine (FMP) or N-formyl methionyl leucylphenylalanine (FMLP)) and concanavalin A. The anisotropy of diphenylhexatriene in macrophages showed a dependence on stimulation both in the rate of decay and in the value of anisotropy at infinite time. Subtle differences were observed between the effect of concanavalin A and FMLP on the membrane lipid fluidity as detected by fluorescence anisotropy. Concanavalin A stimulation of macrophages decreased the value of the anisotropy at infinite times in the range of 0-20 °C and increased the value at 25-40 °C; and at all temperatures it decreased the rate of decay of anisotropy. At temperatures below 25 °C, the response to FMLP was similar to concanavalin A, but above 25 °C, FMLP only slightly modified the anisotropy decay profile. Another physical parameter, calcium permeability, was examined because Ca+2 fluxes are dependent upon membrane properties. The temperature-dependent profiles of concanavalin A and FMP-stimulated 45Ca+2 efflux from alveolar macrophages were similar. The rate and extent of 45Ca+2 efflux increased from 4 to 22 °C, with no further increases observed up to 37 °C. This pattern correlated well with observed changes in membrane fluidity.

Original languageEnglish
Pages (from-to)299-304
Number of pages6
JournalArchives of Biochemistry and Biophysics
Volume214
Issue number1
DOIs
StatePublished - Mar 1982

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