Scale-associated glycoproteins of Scherffelia dubia (Chlorophyta) form high-molecular-weight complexes between the scale layers and the flagellar membrane
1996
Becker, B. | Perasso, L. | Kammann, A. | Salzburg, M. | Melkonian, M.
Flagellar scales were isolated from the flagellate green alga Scherffelia dubia. The flagellar scales consist mainly of acidic polysaccharides (70%) and glycoproteins (10%), and monosaccharide analyses show that the scales contain high amounts of unusual 2-keto-sugar acids. Approximately, 72 mol % of total carbohydrate is 3-deoxy-manno-2-octulosonic acid, 3-deoxy-5-O-methylmanno-2-octulosonic acid and 3-deoxy-lyxo-2-heptulosaric acid. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed the presence of at least 18 different scale-associated proteins (SAPs), ranging in apparent molecular mass from 77 kDa to over 300 kDa. Lectin blot analyses performed in combination with glycosidase treatment, showed that SAPs contained N-glycans of the high-mannose type and the hybrid type, as well as a complex type that was not immunologically related to higher-plant complex glycans. Most of the SAPs were present in two or possibly three high-molecular-weight complexes. In these complexes, individual polypeptides are cross-linked by disulfide bridges. A polyclonal antibody was raised against a SAP of 126 kDa (SAP126), a glycoprotein present in a high-molecular-weight complex. The SAP126 antibody was used to localize the protein between scale layer and flagellar membrane. We suggest that these high-molecular-weight complexes link scales to the flagellar membrane.
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