The cell adhesion molecule retina cognin is a cell surface protein disulfide isomerase that uses disulfide exchange activity to modulate cell adhesion.

The retina cell adhesion molecule, R-cognin, shares cDNA sequence with protein disulfide isomerase (PDI) but has a different molecular size and subcellular location. We asked whether R-cognin originated from a unique PDI gene transcript or was a product of posttranscriptional processing. The 3'-terminal partial cDNA clone for R-cognin was extended by both 5' RACE and by PCR from sequence near the 5' end of the PDI-translated region. The cDNA sequence was compared to those of chicken, bovine, and human PDI. The R-cognin cDNA sequence was identical to that of chicken PDI and differed by less than 10% from mammalian PDI proteins. The role of the disulfide exchange activity characteristic of both proteins was studied by assessing the cell-aggregation-enhancing ability and tissue specificity of R-cognin and recombinant human PDI and its derivatives. Chicken and normal human PDI proteins showed tissue- and developmental-specific enhancement of cell aggregation identical to R-cognin, and this activity was blocked by inactivation of the -WCGHC- motifs which function in disulfide exchange. Dependence of retina cell aggregation on disulfide exchange activity was shown by blocking that activity with the inhibitor, DTNB, or with a recombinant human PDI with the -WCGHC- motif cysteines mutated. The results suggest that one -WCGHC- motif in R-cognin is sufficient and that the more C-terminal motif is most active. We conclude that R-cognin is a tissue-specific protein product of the standard PDI chicken gene. The -WCGHC- motif in mature R-cognin is necessary, but not sufficient, for cell adhesion.

Thioreductase activity of retina cognin and its role in cell adhesion.

Retina cognin (R-cognin) is a 50-kDa protein on the surface of embryonic chick retina cells that mediates cell-cell recognition and neuronal differentiation. It is developmental stage- and tissue-specific in its expression. The partial cDNA clone for R-cognin is nearly identical to that of chicken protein disulfide isomerase (chicken PDI) and enzyme with thioreductase activity. The R-cognin clone extends from beyond the 3' polyadenylation site up to the boundary between PDI exons 1 and 2, with the putative R-cognin equivalent of PDI exon 1 remaining uncloned. The question posed here was whether the sequence-specific properties of PDI were significant in the action of R-cognin. We show that R-cognin, like PDI, has thioreductase activity as revealed by RNase renaturation enzymatic assays. We then asked if this thioreductase activity was involved in the mediation of cell adhesion and recognition in developing chick retina. We show, through cell aggregation assays, that both R-cognin and chicken PDI enhance chick retina cell aggregation but not that of cells from other CNS tissues. We also show that treating R-cognin and chicken PDI with the thioreductase inhibitor 5,5'-dithio-bis (2-nitrobenzoic acid), which covalently binds to the functional cysteines of the thioreductase active sites, reduces the enhancement of cell aggregation. Thus R-cognin acts, in part, by catalyzing a covalent protein-protein linkage at the cell surface.