Purification of marginal plates from bovine renal peroxisomes: identification with L-alpha-hydroxyacid oxidase B.
The matrix of mammalian peroxisomes frequently contains crystalline inclusions. The most common inclusions are membrane associated plate-like "marginal plates" of hitherto unknown nature in renal peroxisomes and central polytubular "cores" composed of urate oxidase in hepatic peroxisomes. In bovine kidney, peroxisomes of proximal tubules exhibit peculiar angular shapes that are caused by multiple marginal plates (Zaar, K., and H.D. Fahimi. 1990. Cell Tissue Res. 260:409-414). Enriched or highly purified peroxisome preparations from this source were used to purify and characterize marginal plates. By SDS-PAGE, one major polypeptide of Mr 33,500 was observed that corresponded to the marginal plate protein. This polypeptide was identified by its enzymatic activity as well as by immunoblotting and preembedding immunocytochemistry as the isozyme B of L-alpha-hydroxyacid oxidase (EC 1.4.3.2). Morphologically, marginal plates were revealed to consist of rectangular straight-edged sheets, exhibiting a defined crystalline lattice structure. The sheets apparently are composed of a single layer of protomers which associate laterally to form a plate-like structure. As deduced from the negative staining results and the additional information of the thickness of marginal plates, each protomer seems to consist of eight subunits forming a cube-like array. The tendency of L-alpha-hydroxyacid oxidase B to self-associate in vitro (Philips, D.R., J.A. Duley, D.J. Fennell, and R.S. Holmes. 1976. Biochim. Biophys. Acta. 427:679-687) corresponds to the mode of association of cubical protomers to form the so-called marginal plates in renal peroxisomes.
Zaar
K
K
Department of Anatomy and Cell Biology, University of Heidelberg, Germany.
Völkl
A
A
Fahimi
H D
HD
D000429:Alcohol Oxidoreductases / Q000302:isolation & purification*; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002458:Cell Fractionation / Q000379:methods; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007672:Kidney Cortex / Q000648:ultrastructure; D007687:Kidney Tubules, Proximal / Q000648:ultrastructure; D008830:Microbodies / Q000201:enzymology / Q000648:ultrastructure*; D008854:Microscopy, Electron
D000429:Alcohol Oxidoreductases; C021380:L-2-hydroxyacid oxidase