Electron cytochemical demonstration of phosphatase activity with microbody membranes of Basidiobolus haptosporus.
Growth of cells of the potentially zoopathogenic fungus Basidiobolus haptosporus on a nutritionally defined medium with xanthine or urate as the nitrogen source results in greatly increased populations of microbodies. Modified Gomori procedures at the electron microscopic level suggested the single limiting membrane (and in some cases the granular matrix) of immature microbodies to be the exclusive subcellular locale(s) of alkaline phosphatase, 5'-nucleotidase and nucleoside diphosphatase activities. When grown in the presence of low inorganic phosphate, additional alkaline phosphatase activity was further identified cytochemically at and along profiles of endoplasmic reticulum and on inclusions previously described as "double-membraned vesicles". Cytochemical localization of acid phosphatase at microbody membranes was minimal if not ambiguous; Mg++-dependent adenosine triphosphatase and glucose-6-phosphatase were not identified at these locales. Quantitative biochemical estimates of alkaline phosphatase activity levels in particulate fractions initially increased with age of cells, perhaps as a function of the cultural induction and marked increase in immature microbody populations. We suggest that this enzyme may participate in some manner with protein translocation mechanisms associated with microbody biogenesis, ontogeny, and/or physiological function.
Garrison
R G
RG
Research Service, Veterans Administration Medical Center, Kansas City, MO 64128.
Fiskin
A M
AM
D015720:5'-Nucleotidase; D017766:Acid Anhydride Hydrolases; D000469:Alkaline Phosphatase / Q000032:analysis; D001669:Biochemical Phenomena; D001671:Biochemistry; D003470:Culture Media; D004773:Entomophthora / Q000201:enzymology* / Q000254:growth & development / Q000648:ultrastructure; D005658:Fungi / Q000201:enzymology*; D006651:Histocytochemistry; D007425:Intracellular Membranes / Q000201:enzymology; D008830:Microbodies / Q000201:enzymology*; D008854:Microscopy, Electron; D009708:Nucleotidases / Q000032:analysis; D010744:Phosphoric Monoester Hydrolases / Q000032:analysis*
D003470:Culture Media; D009708:Nucleotidases; D000469:Alkaline Phosphatase; D010744:Phosphoric Monoester Hydrolases; D015720:5'-Nucleotidase; D017766:Acid Anhydride Hydrolases; C023837:nucleoside-diphosphatase