Kirrel2 (kin of irregular chiasm-1-like 2), also known as filtrin, NEPH3 (nephrin-like 3) or NLG1 (nephrin-like gene 1), is a 107kD type I transmembrane adhesion protein belonging to the immunoglobulin superfamily.1, 2 Kirrel2 is one of several proteins expressed both in pancreatic islet b cells and kidney glomerulus, and is also found in brain and lymph nodes.1-9 Mouse Kirrel2 cDNA encodes a 700 aa protein that contains a 16 aa signal sequence, a 491 aa extracellular domain with five C2-type Ig-like domains, a 21 aa transmembrane sequence and a 172 aa intracellular domain with a nephrin-like, conserved podocin binding site (mouse aa 589-598).4, 5 An extracellular RGD sequence allows for cell attachment.4 The extracellular domain of mouse Kirrel2 shares 96%, 86%, 85% and 85% amino acid (aa) identity with rat, human, equine and bovine Kirrel2, respectively. It shares 35-37% aa identity with other mouse Kirrel proteins. One mouse isoform that has been described is missing the transmembrane domain (aa 501-533), while another encodes only portions of Ig domains 1-4.5 Human Kirrel2 is downregulated in glomeruli of patients with proteinuria, suggesting an involvement in the maintenance of the glomerular filtration barrier.6 Autoantibodies to Kirrel2 have been identified in a subset of humans with type I diabetes, indicating its involvement with islet b cell function.7 In the brain, Kirrel2 is present in olfactory neurons; but only when they are active; homophilic adhesion of Kirrel2 is postulated to help correlate nerves identifying the same odorant.8 It is also a marker for mouse early postmitotic neural precursors in the ventricular zone of the developing mouse spinal cord.9||A DNA sequence encoding the extracellular domain of mouse Kirrel2 (Met 1-Asp 501; Accession # NP_766486) was fused to a ten-histidine tag at the C-terminus. The protein was expressed in a mouse myeloma cell line, NS0.||Calculated molecular mass of 52.7kD. As a result of glycosylation, the recombinant protein migrates as an ~ 66kD protein in SDS-PAGE under reducing conditions.||Activity: Measured by the ability of the immobilized protein to support the adhesion of MS-1 cells (mouse pancreatic islet endothelial cell line).||Storage and Stability:|Lyophilized powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile PBS and store at -20°C. Reconstituted product is stable for 12 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.

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