Cloning and activity of a novel alpha-latrotoxin from red-back spider venom

Graudins, Andis, Little, Michelle J., Pineda, Sandy S., Hains, Peter G., King, Glenn F., Broady, Kevin W. and Nicholson, Graham M. (2012) Cloning and activity of a novel alpha-latrotoxin from red-back spider venom. Biochemical Pharmacology, 83 1: 170-183. doi:10.1016/j.bcp.2011.09.024

Author Graudins, Andis
Little, Michelle J.
Pineda, Sandy S.
Hains, Peter G.
King, Glenn F.
Broady, Kevin W.
Nicholson, Graham M.
Title Cloning and activity of a novel alpha-latrotoxin from red-back spider venom
Journal name Biochemical Pharmacology   Check publisher's open access policy
ISSN 0006-2952
Publication date 2012-01
Year available 2011
Sub-type Article (original research)
DOI 10.1016/j.bcp.2011.09.024
Volume 83
Issue 1
Start page 170
End page 183
Total pages 14
Place of publication United States
Publisher Elsevier
Collection year 2012
Language eng
Abstract The previous termvenomnext term of the European blackwidow previous termspidernext termLatrodectustredecimguttatus(Theridiidae) contains several high molecular mass (110–140 kDa) neurotoxins that induce neurotransmitter exocytosis. These includea vertebrate-specific α-previous termlatrotoxinnext term (α-LTX-Lt1a) responsible for the clinical symptoms of latrodectism andnumerous insect-specific latroinsectoxins (LITs). In contrast,little is known about the expression of these toxins in other Latrodectus species despite the fact that envenomation by these previous termspidersnext term induces previous termanext term similar clinical syndrome. Here we report highly conserved α-LTX, α-LIT and δ-LIT sequence tagsinL.mactans, L.hesperus and L. hasseltiprevious termvenomsnext term using tandem mass spectrometry,following bioassay-guidedseparation of previous termvenomsnext term by liquid chromatography. Despite this sequence similarity, we show that the anti-α-LTX monoclonal antibody 4C4.1, raised against α-LTX-Lt1a,fails to neutralize the neurotoxicity of all other Latrodectus venomstested in an isolated chick biventer cervicis nerve-muscle bioassay. This suggests that there are important structural differences between α-LTXs in theridiid previous termspidernext termprevious termvenomsnext term. We therefore cloned and sequenced the α-LTX from the Australian previous termrednext term-previous termbacknext termprevious termspidernext termL. hasselti (α-LTX-Lh1a). The deduced amino acid sequence of the mature α-LTX-Lh1acomprises 1180 residues(∼132 kDa) with ∼93%sequenceidentity with α-LTX-Lt1a. α-LTX-Lh1a is composed of an N-terminal domainand previous terma central region containing22 ankyrin-like repeats. The presence of two furin cleavage sites,conserved with α-LTX-Lt1a, indicates thatα-LTX-Lh1ais derived from the proteolytic cleavage of an N-terminal signal peptide and C-terminal propeptide region. However, we show thatα-LTX-Lh1a has key substitutions in the 4C4.1 epitope that explains the lack of binding of the monoclonal antibody.
Keyword Latrodectus
Previous termlatrotoxin
Neurotransmitter release
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2012 Collection
School of Chemistry and Molecular Biosciences
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Citation counts: TR Web of Science Citation Count  Cited 14 times in Thomson Reuters Web of Science Article | Citations
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Created: Fri, 07 Oct 2011, 14:21:03 EST by Professor Glenn King on behalf of School of Chemistry & Molecular Biosciences