Glycoproteins in the matrix of glyoxysomes in endosperm of castor bean seedlings

Abstract

The matrix of glyoxysomes from endosperm of castor bean (Ricinus communis cv Hale) seedlings has been analyzed for the presence of glycosylated proteins. Glyoxysome preparations were monitored for organelle homogeneity by electron microscopy and enzyme marker activities. Glyoxysomes were essentially free of endoplasmic reticulum, mitochondria, and protein bodies. At least eight glyoxysomal matrix glycopeptides ranging in size from 39 to 160 kilodaltons were identified by their affinity for concanavalin A. The glyoxysomal glycoproteins were shown to be radioactively labeled when endosperm was allowed to incorporate glucosamine. Incorporation of glucosamine was inhibited by tunicamycin under conditions which did not inhibit protein synthesis. Hydrolysis of glyoxysomal extracts and subsequent analysis by paper chromatography showed that the labeled precursor was incorporated into the glycoprotein without prior dispersion of the label into amino acids. The present data demonstrate the occurrence of N-linked, high mannose oligosaccharides on polypeptides of the glyoxysomal matrix. This finding is discussed in relation to pathways of protein maturation and transport during glyoxysomal biogenesis.