5'-Nucleotidase, Recombinant, Mouse (CD73, Ecto-5'nucleotidase, 5-NT)

Blue Ice

59

Supplied as a liquid in 20mM Tris, 0.12M sodium chloride, pH 7.5, 4mM CaCl2, 20% glycerol.

CD73 (Ecto-5'nucleotidase, 5′'-NT) is a membrane-bound glycoprotein that functions to hydrolyze extracellular nucleoside monophosphates into bioactive nucleoside intermediates. This enables the uptake of adenosine, inosine, and guanosine into the cell and their subsequent reconversion into ATP and GTP in the purine salvage pathway. The expression of CD73 on neuronal cells has been described during developmental processes and, on lymphocytes, CD73 serves as a maturation marker, being absent from the surface of both immature B and T cells.

Recombinant mouse 5’-Nucleotidase/CD73 ectodomain (rmCD73) encoded by the NT5E gene was expressed with a C-terminal 6X His tag in a Chinese Hamster Ovary cell line. CD73, an ecto-5’-nucleotidase, is an ectoenzyme that is attached to the cell membrane by a glycosyl phophatidylinositol anchor (1, 2). The enzyme is expressed by most cell types. The 5’-nucleotidase activity of CD73 converts extracellular nucleoside-5’-monophosphates to nucleosides. CD73 is one of several enzymes responsible for the production of extracellular adenosine, a signaling molecule that is involved in responses to inflammation and tissue injury (3).

Recombinant protein corresponding to Trp29-Lys549 from mouse 5'-Nucleotidase/CD73, fused to a C-terminal 6X His-Tag, expressed in CHO cells.

The purified, secreted rmCD73 has a N-terminal sequence of W29ELTILHTND. The 527 amino acid rmCD73 predicts a molecular mass of 59kD and migrates as a 70kD band in SDS-PAGE under reducing conditions.

Biological Activity:
Measured by its ability to hydrolyze the 5’-phosphate group from the substrate adenosine-5’-monophosphate (AMP). The orthophosphate product is measured by a Malachite Green Phosphate Detection Kit.

Specific Activity:
~10, 000 pmoles/min/ug.

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.