Recombinant Mouse Pro-EMAP-II Protein, CF

• Reactivity: Mu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Mouse Pro-EMAP-II Protein, CF Summary

• Details of Functionality: Measured by its ability to induce TNF-alpha secretion by RAW 264.7 mouse monocyte/macrophage cells. When rmEMAP-II is coated, it will induce TNF-alpha production by RAW 264.7 cells in the presence of Polymyxin B (20 µg/mL) with an ED₅₀ range of 3-10 µg/mL.
• Source: E. coli-derived mouse EMAP-II protein
  Ala2-Lys310, with an N-terminal Met and 6-His tag
• Accession #: P31230
• N-terminal Sequence: Met
• Protein/Peptide Type: Recombinant Proteins
• Gene: Aimp1
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 34.7 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse Pro-EMAP-II Protein, CF

• AIMP1
• aminoacyl tRNA synthetase complex-interacting multifunctional protein 1
• EMAP-2
• EMAPII
• EMAP-II
• EMAPIImember 1 (endothelialmonocyte-activating)
• endothelial-monocyte activating polypeptide II
• Multisynthase complex auxiliary component p43
• multisynthetase complex auxiliary component p43
• SCYE1

Background

EMAP-II (endothelial monocyte-activating polypeptide II; also AIMP1 and SCYE1) is a 23 kDa polypeptide originally identified in the supernatant of TNF-alpha treated Meth-A fibrosarcoma cells (1, 2). In addition to EMAP-I/S100C and EMAP-III/VEGF, EMAP-II was one of three molecules presumed to enhance TNF-alpha -induced tissue factor expression on endothelial cells. Pro-EMAP-II, otherwise known as p43, is the 310 amino acid (aa) precursor to EMAP-II (3). It is considered to have widespread, if not ubiquitous expression, and is found both intra- and extracellularly (1, 4, 5). Pro-EMAP-II contains no canonical signal sequence. It does possess a 144 aa N-terminus that is characterized by the presence of two coiled-coil regions (aa 6 - 77) and two heparin-binding motifs (aa 71 - 73 and 120 - 122) (6, 7, 8). The C-terminal 166 aa are synonymous with EMAP‑II. This region binds t-RNA (5, 8). Although Pro‑EMAP‑II is 34 kDa in size, it can run anomalously in SDS-page at 43 - 44 kDa (1, 5, 9, 10). When secreted, Pro-EMAP-II circulates as either a dimer or higher-order oligomer, likely due to its coiled-coil region (5). Mouse Pro-EMAP-II shares 89% and 86% aa identity with rat and human Pro-EMAP-II, respectively, and is known to be active on human cells (3, 11). Within the cell, Pro-EMAP-II is one of eleven subunits that constitute the mammalian multisynthetase complex. This is essential for ribosomal protein synthesis (12). When cells are stressed, they dissociate Pro-EMAP-II from this complex and release it (1, 10, 13). Although the particulars are unclear, it would appear that secreted Pro-EMAP-II binds to cell surface ATP synthase alpha -subunits (14). Here, it presumably either acts as a cytokine, or undergoes proteolytic cleavage, first by MMPs at Pro108 (based on human), and later by cathepsin L at Ser145 (10, 15). The cleavage products are monomers and likely not active cytokines (5, 10, 14). In theory, Pro-EMAP-II acts as a mediator of tissue restructuring. Upon cell stress (and subsequent apoptosis), Pro-EMAP-II is released and establishes an environment where phagocytic cells can migrate to, and actively remove, dying cells and cellular debris (1, 5, 10, 13, 16).

1. Van Horssen, R. et al. (2006) Cytokine Growth Factor Rev. 17:339.
2. Kao, J. et al. (1994) J. Biol. Chem. 267:20239.
3. Kao, J. et al. (1994) J. Biol. Chem. 269:25106.
4. Murray, J.C. et al. (2000) Am. J. Pathol. 157:2045.
5. Shalak, V. et al. (2001) J. Biol. Chem. 276:23769.
6. Kim, Y. et al. (2000) J. Biol. Chem. 275:27062.
7. Ahn, H-C. et al. (2003) FEBS Lett. 542:119.
8. Chang, S-Y. et al. (2005) Mol. Pharmacol. 67:1534.
9. Knies, U.E. et al. (1998) Proc. Natl. Acad. Sci. USA 95:12322.
10. Shalak, V. et al. (2007) J. Biol. Chem. 282:10935.
11. Kao, J. et al. (1994) J. Biol. Chem. 269:9774.
12. Robinson J-C. et al. (2000) J. Mol. Biol. 304:983.
13. Ko, Y-G. et al. (2001) J. Biol. Chem. 276:23028.
14. Chang, S.Y. et al. (2002) J. Biol. Chem. 277:8388.
15. Liu, J. and M.A. Schwarz (2006) Exp. Cell Res. 312:2231.
16. Barnett, G. et al. (2000) Cancer Res. 60:2850.

Bioinformatics

• Gene Symbol: Aimp1
• Uniprot:
  • P31230()