Molecular mechanisms for generating transmembrane proton gradients.

Membrane proteins use the energy of light or high energy substrates to build a transmembrane proton gradient through a series of reactions leading to proton release into the lower pH compartment (P-side) and proton uptake from the higher pH compartment (N-side). This review considers how the proton affinity of the substrates, cofactors and amino acids are modified in four proteins to drive proton transfers. Bacterial reaction centers (RCs) and photosystem II (PSII) carry out redox chemistry with the species to be oxidized on the P-side while reduction occurs on the N-side of the membrane. Terminal redox cofactors are used which have pKas that are strongly dependent on their redox state, so that protons are lost on oxidation and gained on reduction. Bacteriorhodopsin is a true proton pump. Light activation triggers trans to cis isomerization of a bound retinal. Strong electrostatic interactions within clusters of amino acids are modified by the conformational changes initiated by retinal motion leading to changes in proton affinity, driving transmembrane proton transfer. Cytochrome c oxidase (CcO) catalyzes the reduction of O2 to water. The protons needed for chemistry are bound from the N-side. The reduction chemistry also drives proton pumping from N- to P-side. Overall, in CcO the uptake of 4 electrons to reduce O2 transports 8 charges across the membrane, with each reduction fully coupled to removal of two protons from the N-side, the delivery of one for chemistry and transport of the other to the P-side.

MCCE analysis of the pKas of introduced buried acids and bases in staphylococcal nuclease.

The pK(a)s of 96 acids and bases introduced into buried sites in the staphylococcal nuclease protein (SNase) were calculated using the multiconformation continuum electrostatics (MCCE) program and the results compared with experimental values. The pK(a)s are obtained by Monte Carlo sampling of coupled side chain protonation and position as a function of pH. The dependence of the results on the protein dielectric constant (ε(prot)) in the continuum electrostatics analysis and on the Lennard-Jones non-electrostatics parameters was evaluated. The pK(a)s of the introduced residues have a clear dependence on ε(prot,) whereas native ionizable residues do not. The native residues have electrostatic interactions with other residues in the protein favoring ionization, which are larger than the desolvation penalty favoring the neutral state. Increasing ε(prot) scales both terms, which for these residues leads to small changes in pK(a). The introduced residues have a larger desolvation penalty and negligible interactions with residues in the protein. For these residues, changing ε(prot) has a large influence on the calculated pK(a). An ε(prot) of 8-10 and a Lennard-Jones scaling of 0.25 is best here. The X-ray crystal structures of the mutated proteins are found to provide somewhat better results than calculations carried out on mutations made in silico. Initial relaxation of the in silico mutations by Gromacs and extensive side chain rotamer sampling within MCCE can significantly improve the match with experiment.

Hyaluronic acid functionalized gold nanorods combined with copper-based therapeutic agents for chemo-photothermal cancer therapy.

We herein report a hybrid nanocomposite (AuNRs-CTN@THA) which is based on hyaluronic acid-coated gold nanorods with loading of a copper complex through strong bonds. AuNRs-CTN@THA exhibits durable photothermal conversion capacity for pH-dominant and pH/temperature dual sensitive drug release, accomplishing synergetic antitumor efficacy and deep tumor penetration.

[Microcosm experiments on the influence of different N/P ratios on phytoplankton community growth in the East China Sea].

In the present study, a microcosm experiment was conducted in situ for 30 days, in order to investigate the effects of different N/P ratios (1N: 1P, 4N: 1P, 8N: 1P, 16N: 1P, 32N: 1P, 64N: 1P, 128N: 1P, and 256N: 1P) on phytoplankton community growth in the East China Sea. The results indicated that the species number, cell abundance, Chl-a content, specific growth rate as well as species composition of the phytoplankton community significantly varied with the N/P ratios. After 6 days of culture, the species number, cell abundance, Chl-a content and specific growth rate in the high N/P ratio groups were significantly higher than those in the low N/P ratio groups. After 30 days of culture, the cell abundance in the groups close to the Redfield ratio (8N: 1P, 16N: 1P, and 32N: 1P) was significantly higher than those in the other treatments. On the other hand, the phytoplankton community in all the treatments was observed a definite succession from diatoms to dinoflagellates during the present study. Nonetheless, the N/P ratios strongly affected the duration of dinoflagellate bloom as well as the dominated species: at the beginning of culture (0th day-12th day), the diatoms dominated the phytoplankton community; then the relative contribution of dinoflagellates to the total cell abundance exceeded diatoms in the 4N: 1P, 16N: 1P, and 32N: 1P groups on the 18th day. After that, the relative contribution of dinoflagellates exceeded diatoms ordinally in the other groups from the 24th day to 30th day. At the end of culture, the phytoplankton community was dominated by the dinoflagellates in all the groups except for the 8N: 1P, 16N: 1P, and 32N: 1P treatments.

[Spatial distribution patterns of heterotrophic, nitrogen, and phosphate bacteria in hypoxic zone of Yangtze River Estuary].

In August 15-28, 2009, a preliminary study was conducted on the spatial distribution characteristics of heterotrophic bacteria (HB), inorganic phosphate bacteria (IPB), organic phosphate bacteria (OPB), denitrifying bacteria (DB), and ammonifying bacteria (AB) in the hypoxic zone of Yangtze River Estuary. In the water surface, water bottom, and sediment surface of the zone, the average quantity of AB was the largest, being 307.52 x 10(4) cells x L(-1), 184.50 x 10(4) cells x L(-1), 199.97 x 10(2) cells x g(-1), followed by that of HB (87.35 x 10(4) cfu x L(-1), 86.85 x 10(4) cfu x L(-1), and 19.56 x 10(2) cfu x g(-1)), and of OPB (19.26 x 10(4) cfu x L(-1), 18.82 x 10(4) cfu x L(-1), and 19.56 x10(2) cfu x g(-1), respectively). IPB was only observed within the Yangtze Estuary, south passage of the Estuary, and Zhoushan inshore, and its average quantity in the water surface, water bottom, and sediment surface was 18.50 x 10(4) cfu x L(-1), 31.00 x 10(4) cfu x L(-1), and 7.17 x 10(2) cfu x g(-1) respectively. DB had a wide distribution, but its average quantity was low, being 3.94 x 10(4) cells x L(-1), 23.08 x 10(4) cells x L(-1), and 6.22 x 10(2) cells x g(-1) in the water surface, water bottom, and sediment surface, respectively. Salinity, NO3(-)-N, PO4(3-)-P, SiO3(2)-Si, and pH were the main factors affecting the distribution of HB, IPB, OPB, and DB in water body and sediment surface. The HB, IPB, and OPB in water bottom and sediment surface had significant positive correlation with water temperature; the HB and OPB in water bottom and the IPB in sediment surface were significantly positively correlated with dissolved oxygen (DO); while the AB in sediment surface was significantly negatively correlated with DO. Cluster analysis showed that hypoxia affected the bacterial community structure in sediment surface.