title: Glycerol formation after the breaking of dormancy of Phycomyces blakesleeanus spores. Role of an interconvertible glycerol-3-phosphatase.
abstract: The breaking of dormancy of Phycomyces blakesleeanus spores by a heat shock was followed by a transient production of glycerol, which culminated within 5-10 min and was terminated at 20 min. Extracts of spores contained a magnesium-dependent glycerol-3-phosphatase active on both L-glycerol 3-phosphate and dihydroxyacetone phosphate but having more affinity for the first substrate than for the second. In extracts from dormant spores, the phosphatase was profoundly inhibited by physiological concentrations of inorganic phosphate, which induced cooperativity for the substrate, whereas the enzyme from heat-activated spores was much less inhibited and this difference in kinetic properties persisted after gel filtration of the enzymic preparation. When measured at 1 mM phosphate and 0.1 mM glycerol 3-phosphate, the phosphatase activity was undetectable in dormant spores, increased sharply during the heat treatment and the following 5 min at 25 degrees C, then fell again to a low value by 20 min. A similar transient activation of the enzyme was observed following the breaking of dormancy by incubation of the spores in the presence of 0.1 M ammonium acetate. Incubation of a cell-free extract or of the partially purified glycerol-3-phosphatase in the presence of ATP-Mg and the catalytic subunit of cyclic-AMP-dependent protein kinase released the enzyme from inhibition by phosphate and endowed it with the same kinetic properties as did the heat treatment of the spores. It appears therefore most likely that phosphorylation of glycerol-3-phosphatase by cyclic-AMP-dependent protein kinase causes its activation and that this transient process explains the equally transient formation of glycerol by the spores after the heat shock.
authors.forename: E
authors.initials: E
authors.lastname: van Schaftingen
authors.forename: A J
authors.initials: AJ
authors.lastname: van Laere
mesh_terms: D000242:Cyclic AMP / Q000494:pharmacology; D004789:Enzyme Activation; D005658:Fungi / Q000378:metabolism*; D005990:Glycerol / Q000096:biosynthesis*; D006358:Hot Temperature; D010710:Phosphates / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000502:physiology*; D010800:Phycomyces / Q000378:metabolism*; D011312:Pressure; D011494:Protein Kinases / Q000378:metabolism; D013172:Spores, Fungal / Q000502:physiology*; D013379:Substrate Specificity
chemical_list: D010710:Phosphates; D000242:Cyclic AMP; D011494:Protein Kinases; D010744:Phosphoric Monoester Hydrolases; C040842:glycerol-1-phosphatase; D005990:Glycerol