Structural Requirements for the Binding of Phenylglycosides to the Surface of Protoplasts

Abstract

A variety of phenylglycosides have been synthesized and tested for binding to the surface of protoplasts from suspension-cultured cells of 'Paul's Scarlet" rose (Rosa sp.). Multivalent phenylglycosides in the form of Yariv antigens (1,3,5,-tri-[p-glycosyloxyphenylazol-2,4,6,-trihy- (4,5,10,16) with lesser amounts present in the cytoplasm (1,19) and at the protoplast surface (4,5,17). Functional roles in cell-cell adhesion, pollen-stigma recognition (3), and water retention have been suggested (2, 7) for these macromolecules, although none ofthese proposed functions have been established. ' A property of AGPs that may be related to their biological function is their affinity for certain phenylglycosides. Investigations of this property (4, 8-10, 12, 23) have generally relied on use of synthetic, multivalent probes of the general structure (1 ,3,5,-tri-[p-glycosyloxyphenylazo]-2,4,6-trihydroxybenzene). These probes are commonly referred to as Yariv antigens (24), and the structure of one such Yariv antigen is shown in Figure 1. This particular Yariv antigen, (l-D-Glc)3, contains three l-Dglucoside arms, but similar molecules containing other sugars in either a-or ,3-anomeric linkage can be synthesized (24). Because Yariv antigens self-associate in aqueous solutions to form complexes of 10 to 50 molecules (22), the effective valency of these molecules is probably much greater than the trivalency suggested by their chemical formula.Certain Yariv antigens form tight complexes with AGPs, as evidenced by the facts that these Yariv antigens can precipitate AGPs from aqueous solution (1, 10) and can specifically stain AGPs in isoelectrofocusing gels (17). In a pioneering study, Jermyn and Yeow (10) found that Yariv antigens could precipitate macromolecules from buffer extracts of a wide variety of plants if the Yariv antigens contained ,-linked D-gluco-, Dgalacto-, D-xylo-, malto-, lacto-, or cellobiopyranosyl residues. Since Jermyn and Yeow (10) found that precipitation did not occur with Yariv antigens containing any of these sugars in alinkage, they gave the name 'all ,B-lectins' to the precipitated macromolecules. These ,B-lectins found by Jermyn and Yeow are now known as AGPs (2, 7).The presence of some form of AGP on the plasma membrane was first suggested by observation that Yariv antigens, when used as histochemical reagents, stained the membrane-cell wall interface (4, 5). Larkin (11,12)

Oxidative cross‐linking of plasma membrane arabinogalactan proteins

“…The AGP artificial antigen ~-glucosyl Yariv reagent (1,3,5-Tris [4-~-o-glucopyranosyl-oxyphenylazo]-2,4,6-trihydroxybenzene;Nothnagel and Lyon, 1986;Yariv et aL, 1962) was from Biosupplies Australia Ltd (Parkville, Australia). The anti-AGP mAbs were from immunizations with different kinds of plant cell preparations and extracts (Bradley et aL, 1988;Dewey et aL, 1997;Knox et al, 1989;Norman etaL, 1986;Pennell etal., 1989).…”

Heterogeneity of Arabinogalactan-Proteins on the Plasma Membrane of Rose Cells

“…were grown as described by Nothnagel and Lyon (1986). Seven days after transfer, cells were collected and thoroughly washed with distilled water on a nylon net with 64-pm openings.…”

“…Furthermore, we found that treatment of Rosa cells with (P-D-Glc),, which specifically binds to AGPs, had a strongly inhibitory but reversible effect on cell proliferation (Serpe and Nothnagel, 1994). Most of the (P-D-GIc), was bound to the cell wall in the division-inhibited cells, although plasma membrane AGPs in Rosa also bind (P-D-Gk), (Nothnagel and Lyon, 1986;Komalavilas et al, 1991).…”

β-Galactosyl Yariv Reagent Binds to the β-1,3-Galactan of Arabinogalactan Proteins

“…The interaction of Yariv phenylglycosides with AGP depends on the glycosyl residues attached to the phenylazotrihydroxybenzene core. In particular, b-glucosyl Yariv phenylglycoside (b-Glc-Yariv) and b-galactosyl Yariv phenylglycoside (b-Gal-Yariv) bind to AGPs, whereas a-glucosyl Yariv and a-galactosyl Yariv (a-Gal-Yariv) do not bind to AGPs (Jermyn and Yeow, 1975;Larkin, 1977Larkin, , 1978Nothnagel and Lyon, 1986). Because of the specific interaction with the b-glycosyl Yariv phenylglycosides (b-Yarivs), AGPs were formerly called "b-lectins" (Jermyn and Yeow, 1975;Gleeson and Jermyn, 1979;Nothnagel and Lyon, 1986).…”

“…In particular, b-glucosyl Yariv phenylglycoside (b-Glc-Yariv) and b-galactosyl Yariv phenylglycoside (b-Gal-Yariv) bind to AGPs, whereas a-glucosyl Yariv and a-galactosyl Yariv (a-Gal-Yariv) do not bind to AGPs (Jermyn and Yeow, 1975;Larkin, 1977Larkin, , 1978Nothnagel and Lyon, 1986). Because of the specific interaction with the b-glycosyl Yariv phenylglycosides (b-Yarivs), AGPs were formerly called "b-lectins" (Jermyn and Yeow, 1975;Gleeson and Jermyn, 1979;Nothnagel and Lyon, 1986).The b-Yarivs are useful tools for staining, detection, and quantification of AGPs. Using b-Glc-Yariv, b-lectins were shown to exist in angiosperm, gymnosperm, fern, moss, and liverwort, illustrating the wide distribution of AGPs in the plant kingdom (Jermyn and Yeow, 1975;Clarke et al, 1978).…”