Enzymatic conversion of glutamate to delta-aminolevulinic acid in soluble extracts of Euglena gracilis.
Glutamate was converted to the chlorophyll and heme precursor delta-aminolevulinic acid in soluble extracts of Euglena gracilis. delta-Aminolevulinic acid-forming activity depended on the presence of native enzyme, glutamate, ATP, Mg2+, NADPH or NADH, and RNA. The requirement for reduced pyridine nucleotide was observed only if, prior to incubation, the enzyme extract was filtered through activated carbon to remove firmly bound reductant. Dithiothreitol was also required for activity after carbon treatment. delta-Aminolevulinic acid formation was stimulated by RNA from various plant tissues and algal cells, including greening barley leaves and members of the algal groups Chlorophyta (Chlorella vulgaris, Chlamydomonas reinhardtii), Rhodophyta (Cyanidium caldarium), Cyanophyta (Anacystis nidulans, Synechocystis sp. PCC 6803), and Prochlorophyta (Prochlorothrix hollandica), but not by RNA derived from Escherichia coli, yeast, wheat germ, bovine liver, and Methanobacterium thermoautotrophicum. E. coli glutamate-specific tRNA was inhibitory. Several of the RNAs that did not stimulate delta-aminolevulinic acid formation nevertheless became acylated when incubated with glutamate in the presence of Euglena enzyme extract. RNA extracted from nongreen dark-grown wild-type Euglena cells was about half as stimulatory as that from chlorophyllous light-grown cells, and RNA from aplastidic mutant cells stimulated only slightly. delta-Aminolevulinic acid-forming enzyme activity was present in extracts of light-grown wild-type cells, but undetectable in extracts of aplastidic mutant and dark-grown wild-type cells. Gabaculine inhibited delta-aminolevulinic acid formation at submicromolar concentration. Heme inhibited 50% at 25 microM, but protoporphyrin IX, Mg-protoporphyrin IX, and protochlorophyllide inhibited only slightly at this concentration.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000622:Aminolevulinic Acid / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D005056:Euglena gracilis / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D005971:Glutamates / Q000378:metabolism*; D018698:Glutamic Acid; D006863:Hydrogen-Ion Concentration; D007982:Levulinic Acids / Q000378:metabolism*; D008027:Light; D008274:Magnesium / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D012313:RNA / Q000494:pharmacology; D012329:RNA, Bacterial / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D013696:Temperature
D005971:Glutamates; D007982:Levulinic Acids; D012329:RNA, Bacterial; D012346:RNA, Transfer, Amino Acyl; D018698:Glutamic Acid; D009249:NADP; D012313:RNA; D000622:Aminolevulinic Acid; D000255:Adenosine Triphosphate; D008274:Magnesium