Purification and characterization of a membrane-bound protein kinase from spinach thylakoids

A protein kinase was isolated from spinach thylakoid membranes by solubilization with octyl glucoside and cholate. The enzyme was purified to apparent homogeneity by ammonium sulfate precipitation, gel filtration, and sucrose density centrifugation, followed by affinity chromatography on either Affi-Gel blue (yielding denatured enzyme) or on histone cross-linked to Sepharose (yielding active enzyme). Electrophoresis on denaturing polyacrylamide gels, followed by staining with silver, revealed the kinase as a single band corresponding to an apparent molecular mass of 64 kDa. The active enzyme underwent autophosphorylation and could be detected by autoradiography following incubation with [gamma-32P]ATP and Mg2+ ion. The specific phosphotransferase activity of purified kinase was approximately 30 nmol of phosphate min-1 (mg protein)-1 with lysine-rich histone (III-S or V-S) as substrate; casein was phosphorylated at approximately 30% of this rate. The physiological substrate for the kinase is presumed to be light-harvesting chlorophyll a/b protein complex. In solubilized form, this was phosphorylated at approximately 10% of the rate observed with histone III-S as substrate, or 10-100 times slower than the estimated rate of phosphorylation of the light-harvesting complex in situ. Possible reasons for this shortfall are considered. The kinase is proposed as the principal effector of thylakoid protein phosphorylation and associated State transition phenomena.     

Kinetic evidence for involvement of two cytochrome b-563 hemes in photosynthetic electron transport

The effect of 2-(n-heptyl)-4-hydroxyquinoline N-oxide (HQNO) on the kinetics of cytochrome b-563 and cytochrome c² turnovers following single-turnover flashes was measured in isolated heterocysts. Low concentrations of HQNO (below 3 μM) blocked reoxidation of cytochrome b-563, whereas higher concentrations (above 5 μM) resulted in additional inhibition of cytochrome b-563 oxidation and also inhibited reduction of cytochrome b-563 and cytochrome c. Similar effects on cytochrome b-563 reduction and reoxidation were obtained with a combination of 5 μM HQNO and 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (1–7 μM). In HQNO-inhibited heterocysts, cytochrome c reduction following a flash occurred in three phases with half-times of 0.5, 2.8 and 45 ms. The second phase nearly equalled the cytochrome b-563 reduction in half-time and magnitude. In the presence of HQNO, the reoxidation of cytochrome b-563 following two closely spaced actinic flashes displayed biphasic kinetics. The two phases correspond to reoxidation of cytochrome b-563 in which one or both of the cytochrome b-563 hemes in the cytochrome b–f complex are reduced. These results are interpreted in terms of a Q-loop in which HQNO, at low concentrations, blocks the site of rapid cytochrome b-563 reoxidation and at higher concentrations, also inhibits the site of electron donation by plastoquinol to the cytochrome b-f complex.     

On the mechanism and stoichiometry of the oxidation of hydrazine by illuminated chloroplasts

Hydrazine can support a rapid oxygen uptake in illuminated chloroplasts. The oxygen uptake rate is inhibited by 3-(3,4-dichlorophenyl)-1, 1-dimethylurea but is only slightly increased by added methyl viologen, and little H₂O₂ is produced. The pH optimum for hydrazine-dependent oxygen uptake is much higher than that of the Hill reaction. Addition of Mn (II) increases the rate of oxygen uptake in the light and causes the reaction to continue in the dark, the dark rate being dependent on the duration of the preceding light period. Flash yield experiments show that at least six electrons are transferred from hydrazine per flash compared to one electron per flash when water is the electron donor.     

Planktic coccal and trichal green algae of Lake Volvi,Greece

In the course of three years' investigations (1983–1986) of Lake Volvi, N Greece, 62 species and varieties of chlorococcal algae and 6 species of ulotrichal algae (Chlorophyceae) were found. Although chlorophytes form the largest group of the phytoplankton assemblages of the lake, their biomass is relatively small or even negligeable in some periods of the year in comparison with domination cyanophytes, diatoms and cryptophytes. Morphological variation of diagnostic features is described and documented by original drawings in 53 taxa.     

Plate 457. Mutisia Subspinosa Compositae

Mutisia subspinosa Cav. is described and illustrated and its relationships with other members of Mutisia sect. Guariruma (Cass.) Cabrera discussed. Notes on its cultivation are provided along with a historical insight into the collections of Née, made on the Malaspina expedition.     

Cytochrome function in the cyclic electron transport pathway of chloroplasts

Flash excitation of isolated intact chloroplasts promoted absorbance transients corresponding to the electrochromic effect (P-518) and the α-bands of cytochrome b₆ and cytochrome f. Under conditions supporting coupled cyclic electron flow, the oxidation of cytochrome b₆ and the reduction of cytochrome f had relaxation half-times of 15 and 17 ms, respectively. Optimal poising of cyclic electron flow, achieved by addition of 0.1 μM 3-(3,4-dichlorophenyl)-1,1-dimethylurea, increased phosphorylation of endogenous ADP and prolonged these relaxation times. The presence of NH₄Cl, or monensin plus NaCl, decreased the half-times for cytochrome relaxation to approximately 2 ms. Uncouplers also revealed the presence of a slow rise component in the electrochromic absorption shift, with formation half-time of about 2 ms. The inhibitors of cyclic phosphorylation antimycin and 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone abolished the slow rise in the electrochromic shift and prolonged the uncoupled relaxation times of cytochromes b₆ and f by factors of ten or more.These observations indicate that cytochrome b₆, plastoquinone and cytochrome f participate in a coupled electron transport process responsible for cyclic phosphorylation in intact chloroplasts. Estimations of cyclic phosphorylation rates from 40 to 120 μmol ATP/mg chlorophyll per h suggest that this process can provide a substantial fraction of the ATP needed for CO₂ fixation.     

Contribution to the knowledge of the species composition of summer phytoplankton of Lake Hubsugul,Mongolia

The summer phytoplankton of ultraoligotrophic Lake Hubsugul, Mongolia, was studied in 1985–1989. 24 taxa, mostly belonging to the chlorococcal algae, are illustrated and their main diagnostic features discussed. Two new combinations,Monoraphidium pseudomirabile (Kor?.) Hind. etZagor., andOocystella mongolia (Antip.) Hind. etZagor, are created.