Biosynthesis of heme in the vitamin E-deficient rat.
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Recombinant plasmids able to complement the Saccharomyces cerevisiae ole3 mutation were isolated. The nucleotide sequence responsible for complementation was localized to a 3.5-kb region. The level of delta-aminolevulinate (ALV) synthase activity in wild-type cells was six-fold lower than in plasmid-transformed ole3 mutant cells. Certain clones secreted a compound that supported growth of a lawn of adjacent ole3 mutant cells.
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1. The relative rates of synthesis of aminolaevulate and aminoacetone by particles prepared from avian erythrocytes were measured under various conditions. 2. The production of both amino ketones by fresh particles was about three times greater in anaemia caused by phenylhydrazine and acetylphenylhydrazine than in anaemia caused by removal of 20-30ml. of blood. 3. The synthesis of aminolaevulate by freeze-dried particles decreased more than that of aminoacetone in the absence of added pyridoxal phosphate, in the presence of cyanide and of tris buffer, and after preincubation of the erythrocyte particles. Other differences in the rates of synthesis of the two amino ketones were observed after (a) incubation of particles at different temperatures and (b) storage of homogenized freeze-dried particles at different pH values. 4. It is suggested that these differences in the production of the two amino ketones are due to the presence of two amino ketone synthetases or to two or more isoenzymes of aminolaevulate synthetase. 5. The metabolic significance of aminoacetone in erythrocytes is discussed.