Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to induce RANTES secretion by THP‑1 human acute monocytic leukemia cells. The ED 50 for this effect is 0.3-1 µg/mL, in the presence of 10 µg/mL of a cross-linking antibody Mouse Anti-polyHistidine Monoclonal Antibody (Catalog # ).
Source
Mouse myeloma cell line, NS0-derived Met1-Leu145, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
AnalysisArg23
Predicted Molecular Mass
15.4 kDa
SDS-PAGE
15 ‑ 16 kDa, reducing conditions
3138-GN/CF |
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Formulation Lyophilized from a 0.2 μm filtered solution in Sodium Citrate and NaCl. | ||
Reconstitution Reconstitute at 100 μg/mL in sterile PBS. | ||
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. | ||
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Background: Granulysin
Granulysin (formerly NKG5 or Lymphokine LAG-2) is a member of the saposin-like protein (SAPLIP) family of membrane disrupting proteins (1). Granulysin is expressed in granules of natural killer and activated cytotoxic T cells. It exhibits cytolytic activity against intracellular or extracellular microbes and also tumors, either alone or in synergy with perforin (2). Human granulysin has structural similarity and 30 - 40% aa identity to granulysins and NK-lysins of other mammals such as bovine, porcine and canine; similar peptides in rodents have not been identified (1). The 15 kDa unglycosylated protein contains five helical domains; helix 2 and 3 contain 9 arginines and one cysteine critical for activity. Peptides of either helix 2 or 3 will lyse bacteria, while helix 3 is needed to lyse tumor targets (3, 4). One isoform designated 519 uses a different start codon, has no signal peptide sequence and is poorly expressed (5). A post-translationally processed 9 kDa form is present in acidified granules and is less lytic than the 15 kDa form at granule pH (6). IL-15 is necessary and sufficient for granulysin upregulation in CD8 T cells (2). Nanomolar granulysin promotes chemotaxis and increases production of chemokines by monocytic cells, while micromolar local concentrations are needed for lysis (7). Experimental evidence supports the following mechanism for activity against intracellular pathogens (8). First, granulysin forms clusters in lipid rafts due to interaction of positive charges in helices 2-3 with acidic sphingolipids. After endocytosis, early endosomes fuse with phagosomes, probably regulated by small GTPase rab5. Granulysin binds microbial membranes through charge interactions and disrupts them, probably via scissoring actions of granulysin molecules (9, 10).
References:
Clayberger, C. and A.M. Krensky (2003) Curr. Opin. Immunol. 15:560.
Ma, L.L. et al. (2002) J. Immunol. 169:5787.
Wang, Z. et al. (2000) J. Immunol. 165:1486.
Linde, C.M.A. et al. (2005) Infect. Immun. 73:6332.
Yabe, T. et al. (1990) J. Exp. Med. 172:1159.
Hanson, D.A. et al. (1999) Mol. Immunol. 36:413.
Deng, A. et al. (2005) J. Immunol. 174:5243.
Walch, M. et al. (2005) J. Immunol. 174:4220.
Ernst, W.A. et al. (2000) J. Immunol. 165:7102.
Anderson, D.H. et al. (2003) J. Mol. Biol. 325:355.
Entrez Gene IDs:
10578 (Human)
Alternate Names:
D2S69Elymphocyte-activation gene 2; GNLY; Granulysin; LAG2; LAG-2; Lymphokine LAG-2; Lymphokine; NKG5; NKG5519; Protein NKG5; TLA519; TLA519T-cell activation protein 519; T-lymphocyte activation gene 519
