Prochlorococcus is the most numerically abundant photosynthetic organism in the oceans and plays a role in global carbon cycling. Despite its ecological significance and the availability of over a tho...

Nature Communications - RNA-binding proteins play key roles in post-transcriptional gene regulation. Here, Hemm et al. developed RAPDOR, a widely applicable tool based on Jensen-Shannon Distance...

Single-pot solid phase-enhanced sample preparation (SP3) and trifluoroacetic acid workflows open the door to high-throughput quantitative proteomics in cya

Circadian clocks allow organisms to anticipate daily fluctuations in light and temperature, but how this anticipatory role promotes adaptation to d...

Proceedings of the National Academy of Sciences (PNAS), a peer reviewed journal of the National Academy of Sciences (NAS) - an authoritative source of high-impact, original research that broadly spans...

Abstract. Pigments and dyes are widely used in multiple industries. Ketocarotenoids are a highly sought-after group of pigments with a dark-red hue and str

npj Biological Timing and Sleep - The cyanobacterial circadian clock

The advent of oxygenic photosynthesis by ancient cyanobacteria approximately 2.4 billion years ago profoundly transformed Earth’s landscape. Today, cyanobacteria inhabit a vast array of aquatic habita...

Cyanobacteria are major components of biofilms in light-exposed environments, contributing to nutrient cycling, nitrogen fixation and global biogeochemical processes. Although nitrogen-fixing cyanob.....

Wed 20 Aug - Fri 22 [EDT]: 11th International Symposium on Inorganic Carbon Utilization by Aquatic Photosynthetic Organisms (CCM11) will take place at the University of York on August 20-22 later…

Whole-cell catalysis in cyanobacteria allows the transformation of light energy into chemical energy by co-factor recycling and in situ production of oxygen by photosynthesis, requiring only light, CO...

The universally conserved Sec translocon and the YidC/Oxa1-type insertases mediate biogenesis of alpha-helical membrane proteins, but the molecular basis of their cooperation has remained disputed over decades. A recent discovery of a multi-subunit insertase in eukaryotes has raised the question about the architecture of the putative bacterial ortholog SecYEG-YidC and its functional mechanism. Here, we combine cryogenic electron microscopy with cell-free protein synthesis in nanodiscs to visualize biogenesis of the polytopic membrane protein NuoK, the subunit K of NADH-quinone oxidoreductase, that requires both SecYEG and YidC for insertion. We demonstrate that YidC is recruited to the back of the translocon at the late stage of the substrate insertion, in resemblance to the eukaryotic system, and in vivo experiments indicate that the complex assembly is vital for the cells. The nascent chain does not utilize the lateral gate of SecYEG, but enters the lipid membrane at the SecYE-YidC interface, with YidC being the primary insertase. SecYEG-YidC complex promotes folding of the nascent helices at the interface prior their insertion, so the examined cellular pathway follows the fundamental thermodynamic principles of membrane protein folding. Our data provide the first detailed insight on the elusive insertase machinery in the physiologically relevant environment, highlight the importance of the nascent chain for its assembly, and prove the evolutionary conservation of the gate-independent insertion route. ### Competing Interest Statement The authors have declared no competing interest. Deutsche Forschungsgemeinschaft, https://ror.org/018mejw64, Ke1879/3, 267205415 (CRC 1208) European Research Council, https://ror.org/0472cxd90, CRYOTRANSLATION

Current applications of mass spectrometry-based proteomics range from single cell to body fluid analysis that come with very different demands regarding sensitivity or sample throughput. Additionally, the vast molecular complexity of proteomes and the massive dynamic range of protein concentrations in these biological systems require very high-performance chromatographic separations in tandem with the high speed and sensitivity afforded by mass spectrometer. In this study, we focussed on the chromatographic angle and, more specifically, systematically evaluated proteome analysis performance across a wide range of chromatographic flow rates (0.3 – 50 μL/min) and associated column diameters using a Vanquish Neo UHPLC coupled online to a Q Exactive HF-X mass spectrometer. Serial dilutions of HeLa cell line digests were used for benchmarking and total analysis time from injection-to-injection was intentionally fixed at 60 minutes (24 samples per day). The three key messages of the study are that i) all chromatographic flow rates are suitable for high-quality proteome analysis, ii) capLC (1.5 μL/min) is a very robust, sensitive and quantitative alternative to nanoLC for many applications and iii) showcased proteome, phosphoproteome and drug proteome data provide sound empirical guidance for laboratories in selecting appropriate chromatographic flow rates and column diameters for their specific application. ### Competing Interest Statement R.Z. and C.P. are employees of Thermo Fisher Scientific. BK is a non-operational co-founder and shareholder of MSAID. The other authors declare no competing interests. * ABBREVIATIONS : ACN : Acetonitrile CAA : Chloroacetamide capLC : Capillary Liquid Chromatography CSF : Cerebrospinal Fluid CV : Coefficient of Variation DDA : Data-dependent Acquisition DIA : Data-Independent Acquisition DMSO : Dimethyl Sulfoxide DTT : Dithiothreitol EC50 : Effective Concentration to Reduce 50% of Protein Binding to Kinobeads FBS : Fetal Bovine Serum fmol : Femtomole FDR : False Discovery Rate FWHM : Full Width at Half Maximum HeLa : Human Cervical Carcinoma Cell Line HLB : Hydrophilic-Lipophilic Balance HPLC : High-Performance Liquid Chromatography i.d. : Internal Diameter Kdapp : Apparent Dissociation Constant LC : Liquid Chromatography LC-MS/MS : Liquid Chromatography-Tandem Mass Spectrometry µg : Microgram min : Minute µL : Microliter µLC : Microflow Liquid Chromatography mm : Milimeter mM : Milimolar MS : Mass Spectrometry ng : Nanogram nL : Nanoliter nLC : Nanoflow Liquid Chromatography nM : Nanomolar PSM : Peptide-spectrum Match RT : Retention Time SPD : Samples per Day SWATH-MS : Sequential Window Acquisition of All Theoretical Mass Spectra TFA : Trifluoroacetic Acid Å : Angstrom

Postdoctoral scholar in biochemistry of cyanobacterial CO 2 fixation The Blikstad research group is searching for a postdoc to study fundamental principles of CO 2 fixation in cyanobacteria. Our...