The Steady State Chlorophyll a Fluorescence Exhibits in Vivo an Optimum as a Function of Light Intensity which Reflects the Physiological State of the Plant

Modulated (690 and 730 nm), as well as direct chlorophyll (Chl) a fluorescence and changes in the concentration of the oxidized P700 were measured under steady state conditions in leaves of higher plants adapted to different light intensities. All the leaf samples exhibit an optimum curve of steady state fluorescence yield (Fs) versus the light intensity but its position with respect to light intensity varies considerably from one species to another or from one sample to other even in the same plant or within the same leaf sample. However, the optimum level of Fs was always at a moderate light intensity. By using the modulated fluorescence technique, the system with all closed (Flm) or open reaction center (Flo) were measured in steady state conditions. Each experimentally measured fluorescence yield was separated into a fluorescence emission of open (Fopen = Flo,(1—Vs)) and closed (Fclosed = (Flm . Vs)) reaction center (RC) of photosystem II where Vs=(Fs - Flo)/(Flm - Flo) is the function of fraction of closed reaction centers. With increasing light intensity, the fraction of open RC decreased while the fraction of closed RC increased. Maximum quantum efficiency (ΦPo) and actual quantum efficiency (ΦP) decreased by increasing light intensity. An optimum level of Fs was observed, when the fraction of closed reaction centers Vs of each sample was about 0.2 showing a common quenching mechanism which determines the fluorescence properties under steady state condition. This explains the apparent phenomenological contradiction that the fluorescence yield under steady state conditions can increase or decrease upon an increase of actinic ligh

Demonstration of a stabilized pyramidal nitrogen (sp3) in an acyclic system by 1HNMR spectroscopy: A strong repulsive interaction between the lone electron pair and a phenyl ring

236-24

Stereochemical assignment through intramolecular hydrogen bonded stabilized conformation about N-C (aryl) bond: An 1H NMR study

1030-103

Glutamyl-tRNA Reductase of Chlorobium vibrioforme Is a Dissociable Homodimer That Contains One Tightly Bound Heme per Subunit

δ-Aminolevulinic acid, the biosynthetic precursor of tetrapyrroles, is synthesized from glutamate via the tRNA-dependent five-carbon pathway in the green sulfur bacterium Chlorobium vibrioforme. The enzyme glutamyl-tRNA reductase (GTR), encoded by the hemA gene, catalyzes the first committed step in this pathway, which is the reduction of tRNA-bound glutamate to produce glutamate 1-semialdehyde. To characterize the GTR protein, the hemA gene from C. vibrioforme was cloned into expression plasmids that added an N-terminal His(6) tag to the expressed protein. The His-tagged GTR protein was purified using Ni affinity column chromatography. GTR was observable as a 49-kDa band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels. The native molecular mass, as determined by gel filtration chromatography, appeared to be approximately 40 kDa, indicating that native GTR is a monomer. However, when the protein was mixed with 5% (vol/vol) glycerol, the product had an apparent molecular mass of 95 kDa, indicating that the protein is a dimer under these conditions. Purified His(6)-GTR was catalytically active in vitro when it was incubated with Escherichia coli glutamyl-tRNA(Glu) and purified recombinant Chlamydomonas reinhardtii glutamate-1-semialdehyde aminotransferase. The expressed GTR contained 1 mol of tightly bound heme per mol of pep tide subunit. The heme remained bound to the protein throughout purification and was not removed by anion- or cation-exchange column chromatography. However, the bound heme was released during SDS-PAGE if the protein was denatured in the presence of β-mercaptoethanol. Added heme did not inhibit the activity of purified expressed GTR in vitro. However, when the GTR was expressed in the presence of 3-amino-2,3- dihydrobenzoic acid (gabaculine), an inhibitor of heme synthesis, the purified GTR had 60 to 70% less bound heme than control GTR, and it was inhibited by hemin in vitro