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Identification of the binding site for the Lutheran blood group glycoprotein on laminin alpha 5 through expression of chimeric laminin chains in vivo.

The Lutheran blood group glycoprotein (Lu), also known as basal cell adhesion molecule, is an Ig superfamily transmembrane receptor for laminin alpha5. Lu is expressed on the surface of a subset of muscle and epithelial cells in diverse tissues and is thought to be involved in both normal and disease processes, including sickle cell disease and cancer. Here we investigated the binding of Lu to laminin alpha5 in vivo and in vitro. We prepared a soluble recombinant Lu (sol-Lu) composed of the Lu extracellular domain and a His(6) tag. Sol-Lu bound specifically to laminin-10/11 (alpha5beta1/beta2gamma1) in enzyme-linked immunosorbent assays and bound to bona fide basement membranes containing laminin alpha5 in tissue sections. Sol-Lu did not bind to tissue sections of laminin alpha5 knockout embryos, despite the fact that the four other alpha chains were present. To identify the Lu-binding site on laminin alpha5, we prepared modified alpha5 cDNAs encoding chimeric laminins containing all or part of the laminin alpha1 G domain in place of the analogous alpha5 regions. These constructs were used to generate transgenic mice. Proteins derived from transgenes were detected in basement membranes and were assayed for their ability to bind Lu by examining the localization of endogenous Lu and the binding of sol-Lu applied to tissue sections. Our results demonstrate that the alpha5 LG3 module is essential for Lu binding to laminin alpha5.

Pubmed ID: 12244066 RIS Download

Mesh terms: Animals | Basement Membrane | Binding Sites | Cell Adhesion Molecules | DNA, Complementary | Embryo, Mammalian | Heart | Humans | Immunohistochemistry | Laminin | Lutheran Blood-Group System | Mice | Mice, Knockout | Mice, Transgenic | Muscle, Skeletal | Neoplasm Proteins | Protein Binding | Recombinant Fusion Proteins | Transgenes

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Associated grants

  • Agency: NIDDK NIH HHS, Id: P50 DK045181
  • Agency: NIGMS NIH HHS, Id: R01 GM060432

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