Inorganic sulfur oxidizing system in green sulfur bacteria.

Green sulfur bacteria use various reduced sulfur compounds such as sulfide, elemental sulfur, and thiosulfate as electron donors for photoautotrophic growth. This article briefly summarizes what is known about the inorganic sulfur oxidizing systems of these bacteria with emphasis on the biochemical aspects. Enzymes that oxidize sulfide in green sulfur bacteria are membrane-bound sulfide-quinone oxidoreductase, periplasmic (sometimes membrane-bound) flavocytochrome c sulfide dehydrogenase, and monomeric flavocytochrome c (SoxF). Some green sulfur bacteria oxidize thiosulfate by the multienzyme system called either the TOMES (thiosulfate oxidizing multi-enzyme system) or Sox (sulfur oxidizing system) composed of the three periplasmic proteins: SoxB, SoxYZ, and SoxAXK with a soluble small molecule cytochrome c as the electron acceptor. The oxidation of sulfide and thiosulfate by these enzymes in vitro is assumed to yield two electrons and result in the transfer of a sulfur atom to persulfides, which are subsequently transformed to elemental sulfur. The elemental sulfur is temporarily stored in the form of globules attached to the extracellular surface of the outer membranes. The oxidation pathway of elemental sulfur to sulfate is currently unclear, although the participation of several proteins including those of the dissimilatory sulfite reductase system etc. is suggested from comparative genomic analyses.

An overview on chlorophylls and quinones in the photosystem I-type reaction centers.

Minor but key chlorophylls (Chls) and quinones in photosystem (PS) I-type reaction centers (RCs) are overviewed in regard to their molecular structures. In the PS I-type RCs, the prime-type chlorophylls, namely, bacteriochlorophyll (BChl) a' in green sulfur bacteria, BChl g' in heliobacteria, Chl a' in Chl a-type PS I, and Chl d' in Chl d-type PS I, function as the special pairs, either as homodimers, (BChl a')(2) and (BChl g')(2) in anoxygenic organisms, or heterodimers, Chl a/a' and Chl d/d' in oxygenic photosynthesis. Conversions of BChl g to Chl a and Chl a to Chl d take place spontaneously under mild condition in vitro. The primary electron acceptors, A (0), are Chl a-derivatives even in anoxygenic PS I-type RCs. The secondary electron acceptors are naphthoquinones, whereas the side chains may have been modified after the birth of cyanobacteria, leading to succession from menaquinone to phylloquinone in oxygenic PS I.

Biochemical studies of a soxF-encoded monomeric flavoprotein purified from the green sulfur bacterium Chlorobaculum tepidum that stimulates in vitro thiosulfate oxidation.

In the green sulfur bacterium Chlorobaculum tepidum, three sulfur oxidizing enzyme system (Sox) proteins, SoxAXK, SoxYZ, and SoxB (the core TOMES, thiosulfate oxidizing multi-enzyme system) are essential to in vitro thiosulfate oxidation. We purified monomeric flavoprotein SoxF from this bacterium, which had sulfide dehydrogenase activity. SoxF enhanced the thiosulfate oxidation activity of the purified core TOMES with various cytochromes as electron acceptors to different degrees without any change in the affinity for thiosulfate. The apparent reaction rates with 50 microM- C. tepidum cytochrome c-554 were slightly higher than with horse-heart cytochrome c, and the addition of 0.5 microM- SoxF increased the rate by 92%. The rates with 50 microM- horse-heart cytochrome c and 50 muM- horse-heart cytochrome c plus 0.5 muM- cytochrome c-554 were increased by SoxF by 31% and 120% respectively. We conclude that SoxF mediates electron transfer between the components of core TOMES and externally added cytochromes.

The effect of hip flexion angle on muscle elongation of the hip adductor muscles during stretching.

In order to perform effective static stretching of the hip adductor muscles, it is necessary to clarify the position where the muscles are most stretched. However, the effective flexion angle in stretching for each adductor muscle remains unclear. The goal of this study was to investigate the effect of hip flexion angle on muscle elongation of hip adductor muscles during stretching. Sixteen healthy men were recruited for this study. Shear elastic modulus, an index of muscle elongation, of the adductor longus (AL), and both the anterior and posterior adductor magnus (anterior AM) were measured using ultrasonic shear wave elastography at rest (supine position) and at 5 stretching positions (maximal hip abduction at 90°, 60°, 30°, 0°, and -15° hip flexion). For the AL, the shear elastic modulus at rest was significantly lower than that in all stretching positions. However, there was no significant difference among stretching positions. For the anterior AM, there was no significant difference between stretching positions and at rest. For the posterior AM, the shear elastic modulus in 90°, 60°, and 30° hip flexion were significantly higher than that at rest. The shear elastic modulus in 90° hip flexion was significantly higher than that in 60° and 30° hip flexion. Our results suggest that the AL is elongated to the same extent by maximal hip abduction regardless of hip flexion angle, the anterior AM is not elongated regardless of the hip flexion angle; the posterior AM is elongated at all angles except at 0° and -15° hip flexion and is most extended at 90° hip flexion.

SoxAX binding protein, a novel component of the thiosulfate-oxidizing multienzyme system in the green sulfur bacterium Chlorobium tepidum.

From the photosynthetic green sulfur bacterium Chlorobium tepidum (pro synon. Chlorobaculum tepidum), we have purified three factors indispensable for the thiosulfate-dependent reduction of the small, monoheme cytochrome c(554). These are homologues of sulfur-oxidizing (Sox) system factors found in various thiosulfate-oxidizing bacteria. The first factor is SoxYZ that serves as the acceptor for the reaction intermediates. The second factor is monomeric SoxB that is proposed to catalyze the hydrolytic cleavage of sulfate from the SoxYZ-bound oxidized product of thiosulfate. The third factor is the trimeric cytochrome c(551), composed of the monoheme cytochrome SoxA, the monoheme cytochrome SoxX, and the product of the hypothetical open reading frame CT1020. The last three components were expressed separately in Escherichia coli cells and purified to homogeneity. In the presence of the other two Sox factors, the recombinant SoxA and SoxX showed a low but discernible thiosulfate-dependent cytochrome c(554) reduction activity. The further addition of the recombinant CT1020 protein greatly increased the activity, and the total activity was as high as that of the native SoxAX-CT1020 protein complex. The recombinant CT1020 protein participated in the formation of a tight complex with SoxA and SoxX and will be referred to as SAXB (SoxAX binding protein). Homologues of the SAXB gene are found in many strains, comprising roughly about one-third of the thiosulfate-oxidizing bacteria whose sox gene cluster sequences have been deposited so far and ranging over the Chlorobiaciae, Chromatiaceae, Hydrogenophilaceae, Oceanospirillaceae, etc. Each of the deduced SoxA and SoxX proteins of these bacteria constitute groups that are distinct from those found in bacteria that apparently lack SAXB gene homologues.

NB-protein (BchN-BchB) of dark-operative protochlorophyllide reductase is the catalytic component containing oxygen-tolerant Fe-S clusters.

Dark-operative protochlorophyllide (Pchlide) oxidoreductase is a nitrogenase-like enzyme consisting of the two components, L-protein (BchL-dimer) and NB-protein (BchN-BchB-heterotetramer). Here, we show that NB-protein is the catalytic component with Fe-S clusters. NB-protein purified from Rhodobacter capsulatus bound Pchlide that was readily converted to chlorophyllide a upon the addition of L-protein and Mg-ATP. The activity of NB-protein was resistant to the exposure to air. A Pchlide-free form of NB-protein purified from a bchH-lacking mutant showed an absorption spectrum suggesting the presence of Fe-S centers. Together with the Fe and sulfide contents, these findings suggested that NB-protein carries two oxygen-tolerant [4Fe-4S] clusters.

Characterization of cytosolic tetrapyrrole-binding proteins in Arabidopsis thaliana.

In plant cells, tetrapyrroles are synthesized in plastids and distributed to numerous organelles to function in various vital activities. However, molecular mechanisms of tetrapyrroles trafficking in plant cells are poorly understood. In animal cells, experimental evidence suggests that the p22HBP/SOUL family are cytosolic heme carrier proteins functioning in heme trafficking. In this study, we characterized Arabidopsis cytosolic heme-binding proteins (cHBPs) homologous to the p22HBP/SOUL family. Six homologous genes were identified in the complete genome of Arabidopsis. Deduced amino acid sequences of two genes contained N-terminal amino acid extensions, presumably functioning as signal peptides to organelles. No such extension was observed in the other four genes, but one gene contained a ten-base deletion in its open reading frame, suggesting it maybe a pseudogene. The remaining three genes encoding putative cHBPs, designated cHBP1, cHBP2 and cHBP3, were further analyzed. Semiquantitative RT-PCR analysis showed that cHBP1 was preferentially expressed in leaves, while cHBP2 was predominantly expressed in roots. A tetrapyrrole binding assay using recombinant proteins of cHBP1 and cHBP2 revealed that both cHBPs bind to heme, protoporphyrin IX, and Mg-protoporphyrin IX dimethyl ester with distinct dissociation constants (Kd) of approximately submicro molar concentrations. Low temperature electron spin resonance (ESR) spectra showed that both cHBP1 and cHBP2 bind high-spin type heme. When mixed with apo-horse radish peroxidase (HRP), heme-bound cHBP1 and cHBP2 showed comparable abilities for reconstitution of HRP activity, showing that both cHBPs bind heme reversibly. These results suggest that both cHBP1 and cHBP2 have properties suitable for tetrapyrrole carrier proteins and function in distinct organs in plant cells.