Substructure of crystalline peroxisomes in methanol-grown Hansenula polymorpha: evidence for an in vivo crystal of alcohol oxidase.
The substructural organization of completely crystalline peroxisomes present in Hansenula polymorpha cells grown under methanol limitation in a chemostat was investigated by different cytochemical and ultrastructural techniques. Time-dependent cytochemical staining experiments indicated that activities of the two main constituents of these organelles, namely, alcohol oxidase  and catalase, were present throughout the crystalline matrix. Catalase was completely removed from isolated peroxisomes by osmotic shock treatment. After such treatment, the ultrastructure of the crystalline matrix of the organelles remained virtually intact. Because alcohol oxidase activity was still present in this matrix, it was concluded that alcohol oxidase protein is the only structural element of the peroxisomal crystalloids. The molecular architecture of the crystalloids was investigated in ultrathin cryosections which permitted recognition of individual molecules in the crystalline matrix. Depending on the plane of sectioning, different crystalline patterns were observed. Tilting experiments indicated that these images were caused by superposition of octameric alcohol oxidase molecules arranged in a tetragonal lattice. A three-dimensional model of the crystalloid is presented. The repeating unit of this structure is composed of four alcohol oxidase molecules. The crystalloid represents an open structure, which may explain the observed free mobility of catalase molecules.
D000429:Alcohol Oxidoreductases / Q000032:analysis*; D001203:Ascomycota / Q000648:ultrastructure*; D002374:Catalase / Q000032:analysis; D006651:Histocytochemistry; D046911:Macromolecular Substances; D000432:Methanol / Q000378:metabolism; D008830:Microbodies / Q000201:enzymology / Q000648:ultrastructure*; D009940:Organoids / Q000648:ultrastructure*; D010843:Pichia / Q000378:metabolism / Q000648:ultrastructure*; D013104:Spheroplasts / Q000648:ultrastructure
D046911:Macromolecular Substances; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D002374:Catalase; D000432:Methanol