The PufX quinone channel enables the light-harvesting 1 antenna to bind more carotenoids for light collection and photoprotection

Photosynthesis in some phototrophic bacteria requires the PufX component of the reaction centre-light-harvesting 1-PufX (RC-LH1-PufX) complex, which creates a pore for quinone/quinol (Q/QH2 ) exchange across the LH1 barrier surrounding the RC. However, photosynthetic bacteria such as Thermochromatium (T.) tepidum do not require PufX because there are fewer carotenoid binding sites, creating multiple pores in the LH1 ring for Q/QH2 exchange. We show that an αTrp-24 →Phe alteration of the Rhodobacter (Rba.) sphaeroides LH1 antenna impairs carotenoid binding and allows photosynthetic growth in the absence of PufX. We propose that acquisition of PufX and confining Q/QH2 traffic to a pore adjacent to the RC QB site is an evolutionary upgrade that allows increased LH1 carotenoid content for enhanced light absorption and photoprotection. This article is protected by copyright. All rights reserved

Soil radium, soil gas radon and indoor radon empirical relationships to assist in post-closure impact assessment related to near-surface radioactive waste disposal

Least squares (LS), Theil’s (TS) and weighted total least squares (WTLS) regression analysis methods are used to develop empirical relationships between radium in the ground, radon in soil and radon in dwellings to assist in the post-closure assessment of indoor radon related to near-surface radioactive waste disposal at the Low Level Waste Repository in England. The data sets used are (i) estimated 226Ra in the <2 mm fraction of topsoils (eRa226) derived from equivalent uranium (eU) from airborne gamma spectrometry data, (ii) eRa226 derived from measurements of uranium in soil geochemical samples, (iii) soil gas radon and (iv) indoor radon data. For models comparing indoor radon and (i) eRa226 derived from airborne eU data and (ii) soil gas radon data, some of the geological groupings have significant slopes. For these groupings there is reasonable agreement in slope and intercept between the three regression analysis methods (LS, TS and WTLS). Relationships between radon in dwellings and radium in the ground or radon in soil differ depending on the characteristics of the underlying geological units, with more permeable units having steeper slopes and higher indoor radon concentrations for a given radium or soil gas radon concentration in the ground. The regression models comparing indoor radon with soil gas radon have intercepts close to 5 Bq m−3 whilst the intercepts for those comparing indoor radon with eRa226 from airborne eU vary from about 20 Bq m−3 for a moderately permeable geological unit to about 40 Bq m−3 for highly permeable limestone, implying unrealistically high contributions to indoor radon from sources other than the ground. An intercept value of 5 Bq m−3 is assumed as an appropriate mean value for the UK for sources of indoor radon other than radon from the ground, based on examination of UK data. Comparison with published data used to derive an average indoor radon: soil 226Ra ratio shows that whereas the published data are generally clustered with no obvious correlation, the data from this study have substantially different relationships depending largely on the permeability of the underlying geology. Models for the relatively impermeable geological units plot parallel to the average indoor radon: soil 226Ra model but with lower indoor radon: soil 226Ra ratios, whilst the models for the permeable geological units plot parallel to the average indoor radon: soil 226Ra model but with higher than average indoor radon: soil 226Ra ratios

Nanodomains of Cytochrome b(6)f and Photosystem II Complexes in Spinach Grana Thylakoid Membranes

The cytochrome b6f (cytb6f) complex plays a central role in photosynthesis, coupling electron transport between photosystem II (PSII) and photosystem I to the generation of a transmembrane proton gradient used for the biosynthesis of ATP. Photosynthesis relies on rapid shuttling of electrons by plastoquinone (PQ) molecules between PSII and cytb6f complexes in the lipid phase of the thylakoid membrane. Thus, the relative membrane location of these complexes is crucial, yet remains unknown. Here, we exploit the selective binding of the electron transfer protein plastocyanin (Pc) to the lumenal membrane surface of the cytb6f complex using a Pc-functionalized atomic force microscope (AFM) probe to identify the position of cytb6f complexes in grana thylakoid membranes from spinach (Spinacia oleracea). This affinity-mapping AFM method directly correlates membrane surface topography with Pc-cytb6f interactions, allowing us to construct a map of the grana thylakoid membrane that reveals nanodomains of colocalized PSII and cytb6f complexes. We suggest that the close proximity between PSII and cytb6f complexes integrates solar energy conversion and electron transfer by fostering short-range diffusion of PQ in the protein-crowded thylakoid membrane, thereby optimizing photosynthetic efficiency

Five Glutamic Acid Residues in the C-Terminal Domain of the ChlD Subunit Play a Major Role in Conferring Mg2+ Cooperativity upon Magnesium Chelatase

Magnesium chelatase catalyzes the first committed step in chlorophyll biosynthesis by inserting a Mg2+ ion into protoporphyrin IX in an ATP-dependent manner. The cyanobacterial (Synechocystis) and higher-plant chelatases exhibit a complex cooperative response to free magnesium, while the chelatases from Thermosynechococcus elongatus and photosynthetic bacteria do not. To investigate the basis for this cooperativity, we constructed a series of chimeric ChlD proteins using N-terminal, central, and C-terminal domains from Synechocystis and Thermosynechococcus. We show that five glutamic acid residues in the C-terminal domain play a major role in this process

The ChlD subunit links the motor and porphyrin binding subunits of magnesium chelatase

Magnesium chelatase initiates chlorophyll biosynthesis, catalysing the MgATP2- dependent insertion of a Mg2+ ion into protoporphyin IX. The catalytic core of this large enzyme complex consists of three subunits: Bch/ChlI, Bch/ChlD and Bch/ChlH (in bacteriochlorophyll and chlorophyll producing species respectively). The D and I subunits are members of the AAA+ (ATPases associated with various cellular activities) superfamily of enzymes, and they form a complex that binds to H, the site of metal ion insertion. In order to investigate the physical coupling between ChlID and ChlH in vivo and in vitro , ChlD was FLAG-tagged in the cyanobacterium Synechocystis sp. PCC 6803 and co-immunoprecipitation experiments showed interactions with both ChlI and ChlH. Co-production of recombinant ChlD and ChlH in Escherichia coli yielded a ChlDH. Quantitative analysis using microscale thermophoresis (MST) showed magnesium-dependent binding ( K d 331 ± 58 nM) between ChlD and H. The physical basis for a ChlD-H interaction was investigated using chemical crosslinking coupled with mass spectrometry (XL-MS), together with modifications that either truncate ChlD or modify single residues. We found that the C-terminal integrin I domain of ChlD governs association with ChlH, the Mg2+ dependence of which also mediates the cooperative response of the Synechocystis chelatase to magnesium. Our work, showing the interaction site between the AAA+ motor and the chelatase domain of magnesium chelatase, will be essential for understanding how free energy from the hydrolysis of ATP on the AAA+ ChlI subunit is transmitted via the bridging subunit ChlD to the active site on ChlH

Filamentary mass accretion towards the high-mass protobinary system G11.92-0.61 MM2

Funding: S.Z. is funded by the China Scholarship Council-University of St Andrews Scholarship (PhD programmes, No. 201806190010). C.J.C. acknowledges support from the University of St Andrews Restarting Research Funding Scheme (SARRF), which is funded through the SFC grant reference SFC/AN/08/020. J.D.H gratefully acknowledges financial support from the Royal Society (University Research Fellowship; URF\R1\2216.We present deep, sub-arcsecond (∼2000 AU) resolution ALMA 0.82 mm observations of the former high-mass prestellar core candidate G11.92-0.61 MM2, recently shown to be an ~500 AU-separation protobinary. Our observations show that G11.92-0.61 MM2, located in the G11.92-0.61 protocluster, lies on a filamentary structure traced by 0.82 mm continuum and N2H+(4-3) emission. The N2H+(4-3) spectra are multi-peaked, indicative of multiple velocity components along the line of sight. To analyse the gas kinematics, we performed pixel-by-pixel Gaussian decomposition of the N2H+$ spectra using SCOUSEPY and hierarchical clustering of the extracted velocity components using ACORNS. Seventy velocity- and position-coherent clusters (called "trees") are identified in the N2H+-emitting gas, with the 8 largest trees accounting for > 60 per cent  of the fitted velocity components. The primary tree, with ~20 per cent of the fitted velocity components, displays a roughly north-south velocity gradient along the filamentary structure traced by the 0.82 mm continuum. Analysing a ~0.17 pc-long substructure, we interpret its velocity gradient of ~10.5 km s-1pc-1 as tracing filamentary accretion towards MM2 and estimate a mass inflow rate of ~1.8 × 10-4 to 1.2 × 10-3 M⊙ yr-1. Based on the recent detection of a bipolar molecular outflow associated with MM2, accretion onto the protobinary is ongoing, likely fed by the larger-scale filamentary accretion flows. If 50% of the filamentary inflow reaches the protostars, each member of the protobinary would attain a mass of 8 M⊙ within ~1.6 × 105 yr, comparable to the combined timescale of the 70-μm- and mid-infrared-weak phases derived for ATLASGAL-TOP100 massive clumps using chemical clocks.Peer reviewe