title: Nonintegrin laminin receptors in the nervous system: evidence for lack of a relationship to P40.
abstract: Laminins are extracellular matrix proteins that mediate their effects on cells through integrin and nonintegrin receptors. Two receptors of 67 and 110 kD that bind laminin with a high affinity (Kd approximately nM) have been reported in neural cells. Here, we discuss these and other nonintegrin laminin receptors that have been implicated in neural function. In addition, we report studies characterizing a 43 kD protein, (P40), immunologically related to the 67 kD laminin receptor, which may be involved in retinal development. In our studies, polyclonal antisera (anti-P-20-A) to a synthetic peptide derived from the sequence of a cDNA for a putative high-affinity laminin receptor (67 kD) detected a protein of 43 kD in immunoblots of adult rat retinas. Immunohistochemistry with this antiserum showed that the retinal immunoreactivity was predominantly localized in the ganglion cell layer of both adult chicken and rat retinas where it appeared to be intracellular. Retinal ganglion cells were shown to be immunoreactive by retrogradely labeling them from the superior colliculus with a lipophilic dye and subsequently with anti-P-20-A antisera. Consistent with the preferential localization of the P-20-A immunoreactivity in ganglion cells, there was a substantial decrease in the amounts of P40 on Western blots following optic nerve section and resulting retinal ganglion cell death. Screening of a rat (PC12 cell) cDNA library with the anti-P-20-A antiserum further confirmed the specificity of the antiserum for the rat homologue of P40. Rat P40 is 97% identical to the mouse and 87% identical to human P40 at the nucleic acid level and 98% at the protein level. Restriction mapping of the rather abundant positive clones in the library that cross-hybridized with a human cDNA probe for P40 indicated that the full-length cDNA of 1.2 kb was the major and perhaps the only cDNA in the library. In Northern blots of adult rat retina, these clones hybridized to a single 1.2-kb transcript. Electroblots of retinal homogenates probed with radioiodinated laminin demonstrated binding to a broad band at 110 kD, but none at 43 kD. Taken together these findings suggest that P40 may not be a laminin receptor and are in keeping with the hydrophilic composition of the protein, its intracellular localization, as well as other features predicted by its nucleic acid sequence.(ABSTRACT TRUNCATED AT 400 WORDS)
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002645:Chickens / Q000276:immunology / Q000378:metabolism; D004247:DNA / Q000235:genetics; D000939:Epitopes / Q000276:immunology; D005136:Eye Proteins / Q000032:analysis / Q000145:classification* / Q000378:metabolism; D016023:Integrins / Q000276:immunology; D007797:Laminin / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000145:classification* / Q000378:metabolism; D010446:Peptide Fragments / Q000276:immunology; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley / Q000276:immunology / Q000378:metabolism; D017474:Receptors, Laminin / Q000032:analysis / Q000145:classification* / Q000276:immunology / Q000378:metabolism; D012165:Retinal Ganglion Cells / Q000378:metabolism*
chemical_list: D000939:Epitopes; D005136:Eye Proteins; D016023:Integrins; D007797:Laminin; D009419:Nerve Tissue Proteins; D010446:Peptide Fragments; D017474:Receptors, Laminin; D004247:DNA