Certificate of Analysis and Data Sheet
Source:
E.Coli
Catalog No.
PRO-373
Background :
The secreted pore-forming protein listeriolysin O (LLO) is an essential virulence determinant of the facultative intracellular bacterium Listeria monocytogenes that allows the pathogen to escape from the phagocytic vacuole and reach the host cytosol.
LLO is a single polypeptide protein encoded by the hlyA gene and composed
of 529 residues. At its NH2-terminus it possesses a 25 residues long typical signal sequence excited during the secretion process.
Moreover, in its NH2-terminus there is also a 19 amino acids PEST-like sequence that
may target this toxin for degradation. This motif seems to be essential for bacterial virulence too, since its deletion causes a defect in bacterium phagosomal escaping.
Description:
Recombinant Listeriolysin O lacking both the signal secretion sequence
and the PEST- like sequence. LLO-PEST minus is composed of 471 amino acids, starting from amino acid 60 to amino acid 529, with the addition of a methionine in its NH2-terminus.
Physical Appearance:
Sterile Filtered clear solution.
Formulation:
The protein (0.2mg/ml) contains 50mM NaH2PO4, 1mM EDTA, 2.7mM KCl,
1mM DTT, 5% glycerol and 0.5M NaCl.
Stability:
rLLO although stable at 14
0C for 1 week, should be stored desiccated below -18
0C.
Please prevent freeze-thaw cycles.
Please avoid freeze-thaw cycles.
Purity:
Greater than 90.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel
Activity:
1.25x10
6 HU/mg. 2 mM DTT could be use to reactivate the toxin.
Endotoxin:
Less than 0.1ng/ug (IEU/ug) of LLO.
Applications:
a. Cytosolic delivery of molecules, peptides, oligonucleodides and plasmid DNA (4).
b. Production of specific monoclonal antibodies (5).
c. Detection of anti-listeriolysin O antibodies (6).
Usage:
This material is offered by ProSpec-TechnoGene for research,
laboratory or further evaluation purposes.
Latest Publications:
1. Phosphorylation, ubiquitination and degradation of listeriolysin O in mammalian cells: role of the PEST-like sequence.
Cell Microbiol 2006 Feb;8(2):353-64
2. Listeria monocytogenes listeriolysin O and phosphatidylinositol-specific phospholipase C affect adherence to epithelial cells.
Can J Microbiol 2005 Sep;51(9):745-51
3. Detection of Listeria monocytogenes and the toxin listeriolysin O in food.
J Microbiol Methods 2006 Feb;64(2):141-70
4. Multiple synergizing factors contribute to the strength of the CD8+ T cell response against listeriolysin O.
Int Immunol 2006 Jan;18(1):89-100
5. Listeriolysin O, a cytolysin derived from Listeria monocytogenes, inhibits generation of ovalbumin-specific Th2 immune response by skewing maturation of an tigen-specific T cells into Th1 cells.
Clin Exp Immunol 2005 Nov;142(2):268-74
6. Fusion to Listeriolysin O and delivery by Listeria monocytogenes enhances the immunogenicity of HER-2/neu and reveals subdominant epitopes in the FVB/N mouse.
J Immunol 2005 Sep 15;175(6):3663-73