The vacuole-type ATPases (V-ATPases) exist in various intracellular compartments of eukaryotic cells to regulate physiological processes by controlling the acidic environment. The crystal structure of the subunit C of Thermus thermophilus V-ATPase, homologous to eukaryotic subunit d of V-ATPases, has been determined at 1.95-Å resolution and located into the holoenzyme complex structure obtained by single particle analysis as suggested by the results of subunit cross-linking experiments. The result shows that VATPase is substantially longer than the related F-type ATPase, due to the insertion of subunit C between the V 1 (soluble) and the Vo (membrane bound) domains. Subunit C, attached to the Vo domain, seems to have a socket like function in attaching the central-stalk subunits of the V 1 domain. This architecture seems essential for the reversible association͞dissociation of the V 1 and the Vo domains, unique for V-ATPase activity regulation.T he vacuole-type ATPases (V-ATPases) are commonly found in many organisms involved in a variety of physiological processes (1). V-ATPases in eukaryotic cells (eukaryotic VATPases) translocate protons across the membrane consuming ATP. They reside within intracellular compartments, including endosomes, lysosomes, and secretory vesicles, and within plasma membranes of certain cells including renal intercalated cells, osteoclasts, and macrophages. Eukaryotic V-ATPases are responsible for various cell functions including the acidification of intracellular compartments, renal acidification, born resorption, and tumor metastasis (2).V-ATPase and the F-type ATP synthase (F-ATPase) are evolutionary related and share the rotary mechanism coupling ATP synthesis͞hydrolysis and proton translocation across the membrane (2-4). However, these two types of ATPase show significant differences. Reversible association͞dissociation of the V 1 domain (soluble) and the V o domain (membrane bound) is a unique activity regulation mechanism compared to FATPase (Fig. 1). For example, glucose deprivation has been shown to cause a rapid dissociation of the yeast V-ATPase into free V 1 and V o domains, which is reversible and independent of de novo protein synthesis (5, 6). Similar observations have been reported for Manduca sexta and mammalian complexes (7-9). Subunit composition and structure in the stalk region of VATPase, which connects the V o and V 1 domains, are suggested to be significantly different from those in F-ATPase (10) (Fig. 1). Thus, this region is possibly responsible for the association͞ dissociation of the complex.V-ATPases are also found in archaea and some eubacteria (prokaryotic V-ATPases) (11). The V-ATPase from Thermus thermophilus is solely responsible for aerobic ATP synthesis in this bacteria, which lacks F-ATPase (12). The Thermus VATPase is composed of nine different subunits, which are arranged within the atp operon in the order of G-I-L-E-C-F-A-B-D, which encodes proteins with molecular sizes of 13, 71,8,20,35,12, 64, 54, and 25 kDa, respectively (10) (Fig. 1). This A...

PhotosystemIH reaction centers have been studied by femtoend trent absorption spectroseopy. We demonstrate that it is possible to achieve good photoselectivity between the primary electron donor P680 and the majority of the accessory chlorins. Energy tanser can be observed in both directions between P680 and these accessory chlorins depending on which Is initily excited. This oxidizing potential is used to drive water splitting, which gives rise to oxygen evolution. The primary electron donor of PSII is thought to correspond to a spectral feature at 680 nm and is referred to as P680 (3), while a pheophytin (Ph) molecule functions as an electron acceptor (4-6). Studies of PSII core complexes binding 60 and 80 antenna chlorophylls have suggested that the primary radical pair P680+Ph-is formed at a rate of 4100 psfollowing the absorption ofa photon by antenna pigments (7).A similar conclusion was reached from photovoltage studies of larger PSII complexes (8). A kinetic model, in which there is a rapid (--1 ps) equilibration of excitation energy between the antenna pigments and P680, followed by the observed trapping of the excitation energy by radical pair formation in ""100 ps, has been proposed for this process (refs. 9 and 10; reviewed in ref. 2). This so-called trapping limited model (11) is valid when the rate of electron transfer from the primary electron donor is slower than energy transfer back to the antenna pigments. This model has also been applied to other photosynthetic antenna/reaction center complexes (12)(13)(14).However, previous studies have not been able to time resolve the energy-transfer processes that are predicted to cause the equilibration of excitation energy between the antenna and primary electron donor pigments prior to radical pair formation.We report here a study ofexcitation energy equilibration in the isolated Dl/D2/cytochrome b559 complex, which is the reaction center of PSII. This complex is much smaller than the isolated PSII core complexes discussed above, binding only six chlorophyll a and two Ph a pigments (15, 16). While several of these pigments are presumably involved in electron-transfer processes, these pigments will also function in an energy-transferring capacity before charge separation. Time-resolved fluorescence studies have determined that at least 94% of the chlorin pigments in our PSI1 reaction center preparation are able to transfer excitation energy to P680, resulting in a near unity quantum yield of the primary radical pair (17,18). In a previous study, we determined that when P680 is directly excited, Ph reduction occurs primarily at a rate of 21 ps-1 in isolated PSII reaction centers at room temperature (4).There have been several discussions of the similarities between the PSII reaction center of higher plants and the reaction center of purple bacteria (19,20). However, these two complexes are likely to be very different in terms oftheir energy-transfer kinetics when isolated from their antenna systems. The lowest SO-S, opticil transition for the pr...

We report new measurements of the parity-violating asymmetry A(PV) in elastic scattering of 3 GeV electrons off hydrogen and 4He targets with approximately 6.0 degrees . The 4He result is A(PV)=(+6.40+/-0.23(stat)+/-0.12(syst))x10(-6). The hydrogen result is A(PV)=(-1.58+/-0.12(stat)+/-0.04(syst))x10(-6). These results significantly improve constraints on the electric and magnetic strange form factors G(E)(s) and G(M)(s). We extract G(E)(s)=0.002+/-0.014+/-0.007 at =0.077 GeV2, and G(E)(s)+0.09G(M)(s)=0.007+/-0.011+/-0.006 at =0.109 GeV2, providing new limits on the role of strange quarks in the nucleon charge and magnetization distributions.

No abstract

A quinone-independent photoreduction of the low potential form of cytochrome b559 has been studied using isolated reaction centers of photosystem II. Under anaerobic conditions, the cytochrome can be fully reduced by exposure to strong illumination without the addition of any redox mediators. Under high light conditions, the extent and rate of the reduction is unaffected by addition of the exogenous electron donor Mn2+ and, during this process, no irreversible damage occurs to the reaction center. However, prolonged illumination in strong light brings about irreversible bleaching of chlorophyll, indicative of photoinhibitory damage. When the cytochrome is fully reduced and excess Mn2+ is present, the effect of moderate light is to facilitate the photoaccumulation of reduced pheophytin. The dark reoxidation of the reduced cytochrome is very slow under anaerobic conditions but significantly speeded up on addition of oxidized 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone. From these results it is suggested that the low potential form of cytochrome b559 can accept electrons directly from reduced pheophytin and in so doing help to protect the reaction center against acceptor side photoinhibition as suggested by Nedbal et al. [Nedbal, J., Samson, G. & Whitmarsh, J. (1992) Proc. Natl. Acad. Sci. USA 89, 7929-7933]. This conclusion has been incorporated into a model that further suggests that in its high potential form the cytochrome primarily acts to protect against donor side photoinhibition due to increased lifetime of highly oxidized species as previously proposed by Thompson and Brudvig [Thompson, L. & Brudvig, G. W. (1988) Biochemistry 27, 6653-6658]. The particular feature of our scheme is that it incorporates reversible interconversion between the two redox forms so as to protect against either type of photoinhibition.