Protein organization in the matrix of leaf peroxisomes. A multi-enzyme complex involved in photorespiratory metabolism.

This report is an investigation on how the compartmentation of peroxisomal metabolism, involved in the photorespiratory cycle, is accomplished. With isolated peroxisomes from spinach leaves the conversion of serine to glycerate, as coupled to the conversion of glycolate to glycine, was measured. Not only with intact but also with osmotically shocked peroxisomes, which had retained the aggregated state of the peroxisomal matrix but lost the integrity of the boundary membrane, the rates of glycerate synthesis were as high as required for the photorespiratory cycle in vivo. With both intact and shocked peroxisomes the intermediates glyoxylate and hydroxypyruvate did not equilibrate with the medium. It appears from these results that the apparent compartmentation of peroxisomal metabolism is not due to the function of the boundary membrane but to the organization of peroxisomal enzymes in multi-enzyme complexes. When glycolate was added to peroxisomes without transamination partners, glyoxylate was released from the peroxisomes while the peroxisomal matrix partially disintegrated. With solubilized peroxisomes a partial reconstitution of functional enzyme complexes was achieved by the addition of poly(ethylene glycol). The function of the apparently very stable peroxisomal multi-enzyme complexes in protecting the cells from the toxic intermediates H2O2 and glyoxylate is discussed.