Annexin A5 binds to lipopolysaccharide and reduces its endotoxin activity

Jacob H. Rand, Xiao Xuan Wu, Elaine Y. Lin, Alexander Griffel, Philip Gialanella, John C. McKitrick

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Annexin A5 (AnxA5) has a high affinity for phosphatidylserine. The protein is widely used to detect apoptotic cells because phosphatidylserine, a phospholipid that is normally present in the inner leaflets of cytoplasmic membranes, becomes translocated to the outer leaflets during programmed cell death. Here we report the novel observation that AnxA5 binds to Gram-negative bacteria via the lipid A domain of lipopolysaccharide (LPS). Binding of AnxA5 to bacteria was measured quantitatively, confirmed by fluorescence microscopy, and found to be inhibited by antibodies against lipid A. AnxA5 also bound to purified dot-blotted LPS and lipid A. Through ellipsometry, we found that the binding of AnxA5 to purified LPS was calcium dependent and rapid and showed a high affinity- characteristics similar to those of AnxA5 binding to phosphatidylserine. Initial functional studies indicated that AnxA5 can affect LPS activities. AnxA5 inhibited LPS-mediated gelation in the Limulus amebocyte lysate assay. Incubation of LPS with the protein reduced the quantity of tumor necrosis factor alpha (TNF-α) released by cultured monocytes compared to that released upon incubation with LPS alone. Initial in vivo experiments indicated that injection of mice with LPS preincubated with AnxA5 produced serum TNF-αlevels lower than those seen after injection of LPS alone. These data demonstrate that AnxA5 binds to LPS and open paths to investigation of the potential biological and therapeutic implications of this interaction.

Original languageEnglish (US)
JournalmBio
Volume3
Issue number2
DOIs
StatePublished - Mar 2012

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Annexins
Annexin A5
Endotoxins
Lipopolysaccharides
Lipid A
Phosphatidylserines
Tumor Necrosis Factor-alpha
Horseshoe Crabs
Injections
Gram-Negative Bacteria
Fluorescence Microscopy
Monocytes
Phospholipids
Proteins
Cell Death
Cell Membrane
Observation
Calcium
Bacteria

ASJC Scopus subject areas

  • Microbiology
  • Virology

Cite this

Annexin A5 binds to lipopolysaccharide and reduces its endotoxin activity. / Rand, Jacob H.; Wu, Xiao Xuan; Lin, Elaine Y.; Griffel, Alexander; Gialanella, Philip; McKitrick, John C.

In: mBio, Vol. 3, No. 2, 03.2012.

Research output: Contribution to journalArticle

Rand, Jacob H. ; Wu, Xiao Xuan ; Lin, Elaine Y. ; Griffel, Alexander ; Gialanella, Philip ; McKitrick, John C. / Annexin A5 binds to lipopolysaccharide and reduces its endotoxin activity. In: mBio. 2012 ; Vol. 3, No. 2.
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