Further studies on the identification of the phagocytosis receptor of rat retinal pigment epithelial cells

We have previously produced a polyclonal antiserum (R(1)S(5)) against a plasma membrane-enriched fraction of rat retinal pigment epithelial (RPE) cells which inhibits the phagocytosis of photoreceptor outer segments (OS) by these cells, This antiserum has now been used to purify a subset of RPE membrane glycoproteins. Using a combination of lectin affinity chromatography, and chromatography on an affinity column made with R(1)S(5)-IgG, we have enriched an RPE membrane extract about 100-fold. This enriched extract contains only 12 components, all of which are glycoproteins, and retains the ability to adsorb out the inhibitory activity of antiserum R(1)S(5). This shows that one or more of these glycoproteins recognizes an inhibitory IgG in R(1)S(5) and suggests that on- or more of these glycoproteins may participate in the phagocytosis of OS by RPE cells, possibly as the phagocytosis receptor. We have performed N-terminal microsequencing of seven of these glycoproteins: four of the seven, with Mrs of 34, 36, 51 and 55 kDa, show no sequence homology to any known proteins. (C) 1996 Academic Press Limited