Identification of a hemolysin from Actinobacillus pleuropneumoniae and characterization of its channel properties in planar phospholipid bilayers

G. Lalonde, T. V. McDonald, P. Gardner, P. D. O'Hanley

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

A proteinaceous hemolysin secreted by strain 4074 of serotype 1 of Actinobacillus pleuropneumoniae was purified by diafiltration and ion exchange chromatographic techniques. The hemolytic activity is associated with a 107-kDa band as assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and confirmed by Western blotting and immunoprecipitation. This hemolysin produces pores in membranes as demonstrated by osmotic protection studies using red blood cells and carbohydrate compounds of various molecular weights. These assays suggest a pore diameter in the order of 2 nm. Phospholipid bilayers composed of 1:1 w/w phosphotidylserine:phosphotidylethanolamine exposed to this toxin display discrete current flow events typical of transmembrane channels and consistent with the interpretation that this toxin acts by forming pores in phospholipid membranes. The linear relationship of current amplitude to holding potential when examined over the -60 to +60 mV range indicates that this pore has a constant mean single channel conductance level of 350-400 pS.

Original languageEnglish (US)
Pages (from-to)13559-13564
Number of pages6
JournalJournal of Biological Chemistry
Volume264
Issue number23
StatePublished - 1989
Externally publishedYes

Fingerprint

Actinobacillus pleuropneumoniae
Hemolysin Proteins
Phospholipids
Membranes
Ion Exchange
Electrophoresis
Immunoprecipitation
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Assays
Ion exchange
Blood
Erythrocytes
Molecular Weight
Western Blotting
Molecular weight
Cells
Carbohydrates
Serogroup
polyacrylamide gels

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of a hemolysin from Actinobacillus pleuropneumoniae and characterization of its channel properties in planar phospholipid bilayers. / Lalonde, G.; McDonald, T. V.; Gardner, P.; O'Hanley, P. D.

In: Journal of Biological Chemistry, Vol. 264, No. 23, 1989, p. 13559-13564.

Research output: Contribution to journalArticle

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