LRPAP, Recombinant, Murine (LDL Receptor-related Protein-Associated Protein 1)

LRPAP (LDL receptor-related protein-associated protein 1, also RAP) is a ubiquitously expressed 39kD molecular chaperone for LDL receptor family proteins. Mature mouse LRPAP is 332 amino acids (aa) in length and secreted into the ER/Golgi of the cell. It shares 77% and 97% aa sequence identity with human and rat LRPAP, respectively. LRPAP contains three approximately 100 aa alpha-helical domains (D1-D3). The D1 domain contains a low affinity binding site for LRP, and the associated D2 and D3 domains bind LRP with high affinity. Domains D2 and D3 interact with each other, while D1 is independent. The majority of LRPAP is localized in the endoplasmic reticulum and Golgi. LRPAP prevents the premature interaction of LRP, LRP2/megalin, and VLDLR with their coexpressed ligands, thereby promoting proper receptor folding and export from the ER. Protonation of conserved histidine residues within the D3 domain induces the separation of LRPAP and LRP in the relatively acidic Golgi. LRPAP, which contains a C-terminal HNEL motif, can then recycle to the ER. A minor amount of LRPAP remains associated with LRP and can modulate receptor activity on the cell surface. Exogenously applied LRPAP competitively inhibits LDL receptor family binding and uptake of activated alpha 2-macroglobulin, apoB100- or apoE-enriched LDL and VLDL particles, cholesteryl esters, and complexes of PAI-1 with either tPA or uPA. DNA sequence encoding the mature mouse LRPAP (Gln 29-Leu 360, Accession # P55302) with a six residue histidine tag at the C-terminus, was expressed in E. coli. , Unit Definition: , Measured by its ability to bind rcombinant mouse VLDLR with an estimated KD of <0.25nM. Storage and Stability: Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 6 months 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.