title: Expression, characterization and ligand specificity of lipocalin-1 interacting membrane receptor (LIMR).
abstract: Human lipocalin-1 interacting membrane receptor (LIMR) was the first lipocalin receptor to be identified, as a specific receptor for lipocalin-1 (Lcn1). Subsequently LIMR has been reported to interact with other ligands as well, notably with the bovine lipocalin β-lactoglobulin (BLG) and with the unrelated secretoglobin uteroglobin (UG). To study the ligand-binding behaviour of this prototypic lipocalin receptor in more detail, a system was developed for the recombinant expression of LIMR in Drosophila Schneider 2 (S2) cells, and for the subsequent solubilization and purification of the protein. The receptor forms dimers or larger oligomers when solubilized in n-dodecyl β-D-maltoside (DDM). The full-length, functional receptor was captured onto a surface plasmon resonance (SPR) chip via an α-V5 antibody, and the binding of various potential ligands was followed in time. In this way, LIMR was shown to be highly specific for Lcn1, binding the lipocalin with low micromolar to high nanomolar affinity. No interactions with any of the other putative ligands could be detected, raising doubts about the physiological relevance of the reported binding of BLG and UG to the receptor.
authors: University of Leeds, Leeds, UK.
authors: Renske W
authors: RW
authors: Hesselink
authors: John B C
authors: JB
authors: Findlay
mesh_terms: D000818:Animals; D002417:Cattle; D002460:Cell Line; D004330:Drosophila / Q000166:cytology; D015870:Gene Expression; D006801:Humans; D008024:Ligands; D054835:Lipocalin 1 / Q000378:metabolism*; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism*; D020349:Surface Plasmon Resonance; D014598:Uteroglobin / Q000378:metabolism
chemical_list: C430233:LMBR1L protein, human; D008024:Ligands; D054835:Lipocalin 1; D011956:Receptors, Cell Surface; D014598:Uteroglobin