Molecular cloning and characterization of human podocalyxin-like protein - Orthologous relationship to rabbit PCLP1 and rat podocalyxin

Human renal cortex and heart cDNA libraries were screened for a human homolog of rabbit PCLP1 using the rabbit PCLP1 cDNA as a probe, Clones spanning 5569 base pairs with an open reading frame coding for a 528-amino acid peptide a ere obtained, The putative peptide contains a potential signal peptide and a single membrane-spanning region. The extracellular domain contains multiple potential sites for N- and O-linked glycosylation and 4 cysteines for potential disulfide bonding similar to rabbit PCLP1, On Northern blot a major transcript is seen at 5.9 kilobases. Antibodies to this protein show a doublet. at 160/165 kDa on Western blots of human glomerular extract and a pattern of intense glomerular staining and vascular endothelial staining on immunofluorescence of human kidney sections. Comparison of the rabbit and human peptide sequences shows a high degree of identity in the transmembrane and intracellular domains (96%) with a lower degree of identity in the extracellular domain (36%). An antibody to the intracellular domain reacted across species (human, rabbit, and rat) and recognized both rabbit PCLP1 and rat podocalyxin. An interspecies Southern blot probed with a cDNA coding for the intracellular domain showed strong hybridization to all vertebrates tested in a pattern suggesting a single copy gene, Wa conclude that this cDNA and putative peptide represent the human homolog of rabbit PCLP1 and rat podocalyxin.