Recombinant Human Granulysin Protein, CF Summary
Met1-Leu145, with a C-terminal 10-His tag
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|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.
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).
- 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.
Citations for Recombinant Human Granulysin Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
Citations: Showing 1 - 3
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Antimicrobial Properties of an Immunomodulator - 15 kDa Human Granulysin
Authors: Hung-Mu Wei
PLoS ONE, 2016;11(6):e0156321.
Species: Bacteria - E. Coli
Sample Types: Whole Cells
Phenotypic and Functional Characterization of NK Cells in Human Immune Response against the Dimorphic Fungus Paracoccidioides brasiliensis.
Authors: Longhi LN, da Silva RM, Fornazim MC, Spago MC, de Oliveira RT, Nowill AE, Blotta MH, Mamoni RL
J. Immunol., 2012;189(2):935-45.
Sample Types: Whole Cells
Granulysin activates antigen-presenting cells through TLR4 and acts as an immune alarmin.
Authors: Tewary P, Yang D, de la Rosa G
Sample Types: Whole Cells
This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?
The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.
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