Analysis of binding and membrane destabilization of phospholipid membranes by surfactant apoprotein B

Rupert Chang, Shlomo Nir, Francis R Poulain

Research output: Contribution to journalArticle

49 Scopus citations

Abstract

To further elucidate the nature of the molecular interactions of surfactant apoprotein B (SP-B) with phospholipid (PL) membranes, we studied the binding of SP-B to PL membranes and the lipid-dependency of its subsequent effects on leakage and fusion of membranes. SP-B binding to membranes was studied by labeling the protein with the fluorophore 7-nitro-2,1,3-benzoxadiazol-4-yl (NBD) and measuring the fluorescence of the labeled protein in the presence of varying amounts of dipalmitoylphosphatidylcholine-egg phosphatidylglycerol (DPPC-eggPG; 7-3). Leakage of contents from liposomes made of DPPC and varying molar fraction of egg phosphatidylcholine (eggPC) or eggPG was assessed by measuring the fluorescence of entrapped water-soluble probes ANTS and DPX. Fusion of membranes was assessed by measuring the fluorescence of membrane-bound NBD-phosphatidylethanolamine (NBD-PE) and rhodamine-PE (RHO-PE). We found that SP-B bound to PL membranes with high affinity and appeared to irreversibly cluster at the membrane surface, leading to graded release of the vesicle contents and eventually fusion of the membranes with increasing protein-lipid ratios. All lipid mixtures tested were susceptible to the membrane disruptive effects of SP-B, but DPPC-eggPG membranes displayed a biphasic response to increasing molar fractions of eggPG, whereas increasing fractions of eggPC elicited a monotonic response. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)254-264
Number of pages11
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1371
Issue number2
DOIs
StatePublished - May 28 1998
Externally publishedYes

Keywords

  • Lipid-protein interaction
  • Pulmonary surfactant
  • SP-B
  • Surfactant apoprotein
  • Tubular myelius

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Biophysics

Fingerprint Dive into the research topics of 'Analysis of binding and membrane destabilization of phospholipid membranes by surfactant apoprotein B'. Together they form a unique fingerprint.

  • Cite this