Daniel W. Watkins
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View article: A bacterial secretosome for regulated envelope biogenesis and quality control?
A bacterial secretosome for regulated envelope biogenesis and quality control? Open
The Gram-negative bacterial envelope is the first line of defence against environmental stress and antibiotics. Therefore, its biogenesis is of considerable fundamental interest, as well as a challenge to address the growing problem of ant…
View article: Author response: Rate-limiting transport of positively charged arginine residues through the Sec-machinery is integral to the mechanism of protein secretion
Author response: Rate-limiting transport of positively charged arginine residues through the Sec-machinery is integral to the mechanism of protein secretion Open
Article Figures and data Abstract Editor's evaluation Introduction Results Discussion Materials and methods Data availability References Decision letter Author response Article and author information Metrics Abstract Transport of proteins …
View article: A bacterial secretosome for regulated envelope biogenesis and quality control
A bacterial secretosome for regulated envelope biogenesis and quality control Open
SUMMARY As the first line of defence against antibiotics, the Gram-negative bacterial envelope and its biogenesis are of considerable interest to the microbiological and biomedical communities. All bacterial proteins are synthesised in the…
View article: Controlling Protein Nanocage Assembly with Hydrostatic Pressure
Controlling Protein Nanocage Assembly with Hydrostatic Pressure Open
Controlling the assembly and disassembly of nanoscale protein cages for the capture and internalization of protein or non-proteinaceous components is fundamentally important to a diverse range of bionanotechnological applications. Here, we…
View article: Refined measurement of SecA-driven protein secretion reveals that translocation is indirectly coupled to ATP turnover
Refined measurement of SecA-driven protein secretion reveals that translocation is indirectly coupled to ATP turnover Open
Significance The transport of proteins across membranes is fundamental to all life. The biological machinery responsible for this process has been known for some time, but exactly how it works is still contested. In this paper, we describe…
View article: Association of Fluorescent Protein Pairs and Its Significant Impact on Fluorescence and Energy Transfer
Association of Fluorescent Protein Pairs and Its Significant Impact on Fluorescence and Energy Transfer Open
Fluorescent proteins (FPs) are commonly used in pairs to monitor dynamic biomolecular events through changes in proximity via distance dependent processes such as Förster resonance energy transfer (FRET). The impact of FP association is as…
View article: Inter-membrane association of the Sec and BAM translocons for bacterial outer-membrane biogenesis
Inter-membrane association of the Sec and BAM translocons for bacterial outer-membrane biogenesis Open
The outer-membrane of Gram-negative bacteria is critical for surface adhesion, pathogenicity, antibiotic resistance and survival. The major constituent – hydrophobic β-barrel O uter- M embrane P roteins (OMPs) – are first secreted across t…
View article: Author response: Inter-membrane association of the Sec and BAM translocons for bacterial outer-membrane biogenesis
Author response: Inter-membrane association of the Sec and BAM translocons for bacterial outer-membrane biogenesis Open
Article Figures and data Abstract Introduction Results Discussion Materials and methods Data availability References Decision letter Author response Article and author information Metrics Abstract The outer-membrane of Gram-negative bacter…
View article: Controlling protein nanocage assembly with hydrostatic pressure
Controlling protein nanocage assembly with hydrostatic pressure Open
Controlling the assembly and disassembly of nanoscale protein cages for the capture and internalisation of protein or non-proteinaceous components is fundamentally important to a diverse range of bionanotechnological applications. Here, we…
View article: Refined measurement of SecA-driven protein transport reveals indirect coupling to ATP turnover
Refined measurement of SecA-driven protein transport reveals indirect coupling to ATP turnover Open
The universally conserved Sec system is the primary method cells utilise to transport proteins across membranes. Until recently, measuring the activity – a prerequisite for understanding how biological systems works – has been limited to d…
View article: The effect of proximity on the function and energy transfer capability of fluorescent protein pairs
The effect of proximity on the function and energy transfer capability of fluorescent protein pairs Open
Fluorescent proteins (FPs) are commonly used in pairs to monitor dynamic biomolecular events through changes in their proximity via distance dependent processes such as Förster resonance energy transfer (FRET). Many FPs have a tendency to …
View article: Energy landscape steering mediates dynamic coupling in ATP-driven protein translocation by the bacterial Sec machinery
Energy landscape steering mediates dynamic coupling in ATP-driven protein translocation by the bacterial Sec machinery Open
The Sec translocon is a highly conserved membrane complex for transport of polypeptides across, or into, lipid bilayers. In bacteria, the core protein-channel SecYEG resides in the inner-membrane, through which secretion is powered by the …
View article: Positive functional synergy of structurally integrated artificial protein dimers assembled by Click chemistry
Positive functional synergy of structurally integrated artificial protein dimers assembled by Click chemistry Open
Construction of artificial higher order protein complexes allows sampling of structural architectures and functional features not accessible by classical monomeric proteins. Here, we combine in silico modelling with expanded genetic code f…
View article: Rate-limiting transport of positive charges through the Sec-machinery is integral to the mechanism of protein transport
Rate-limiting transport of positive charges through the Sec-machinery is integral to the mechanism of protein transport Open
Transport of proteins across and into membranes is a fundamental biological process with the vast majority being conducted by the ubiquitous Sec machinery. In bacteria, this is usually achieved when the SecY-complex engages the cytosolic A…
View article: Inter-membrane association of the Sec and BAM translocons for bacterial outer-membrane biogenesis
Inter-membrane association of the Sec and BAM translocons for bacterial outer-membrane biogenesis Open
SUMMARY The outer-membrane of Gram-negative bacteria is critical for surface adhesion, pathogenicity, antibiotic resistance and survival. The major constituent – hydrophobic β-barrel O uter- M embrane P roteins (OMPs) – are secreted across…
View article: A High-Resolution Luminescent Assay for Rapid and Continuous Monitoring of Protein Translocation across Biological Membranes
A High-Resolution Luminescent Assay for Rapid and Continuous Monitoring of Protein Translocation across Biological Membranes Open
Protein translocation is a fundamental process in biology. Major gaps in our understanding of this process arise due the poor sensitivity, low time resolution and irreproducibility of translocation assays. To address this, we applied NanoL…
View article: Positive Functional Synergy of Structurally Integrated, Designed Artificial Protein Dimers Assembled by Fully Genetically Encoded Click Chemistry
Positive Functional Synergy of Structurally Integrated, Designed Artificial Protein Dimers Assembled by Fully Genetically Encoded Click Chemistry Open
We combined in silicomodelling with fully genetically encoded strain promoted azide-alkyne cycloaddition, to construct bespoke protein dimers. Using fluorescent proteins GFP and Venus as models, homo and heterodimers were constructed that …
View article: Positive Functional Synergy of Structurally Integrated, Designed Artificial Protein Dimers Assembled by Fully Genetically Encoded Click Chemistry
Positive Functional Synergy of Structurally Integrated, Designed Artificial Protein Dimers Assembled by Fully Genetically Encoded Click Chemistry Open
We combined in silico modelling with fully genetically encoded strain promoted azide-alkyne cycloaddition, to construct bespoke protein dimers. Using fluorescent proteins GFP and Venus as models, homo and heterodimers were constructed that…
View article: A High-Resolution Luminescent Assay for Rapid and Continuous Monitoring of Protein Translocation Across Biological Membranes
A High-Resolution Luminescent Assay for Rapid and Continuous Monitoring of Protein Translocation Across Biological Membranes Open
Protein translocation is a fundamental process in biology. Major gaps in our understanding of this process arises due the poor sensitivity, low time-resolution and irreproducibility of translocation assays. To address this, we applied Nano…
View article: Specific cardiolipin–SecY interactions are required for proton-motive force stimulation of protein secretion
Specific cardiolipin–SecY interactions are required for proton-motive force stimulation of protein secretion Open
Significance Many proteins are located in lipid membranes surrounding cells and subcellular organelles. The membrane can impart important structural and functional effects on the protein, making understanding of this interaction critical. …
View article: Dynamic action of the Sec machinery during initiation, protein translocation and termination
Dynamic action of the Sec machinery during initiation, protein translocation and termination Open
Protein translocation across cell membranes is a ubiquitous process required for protein secretion and membrane protein insertion. In bacteria, this is mostly mediated by the conserved SecYEG complex, driven through rounds of ATP hydrolysi…
View article: Dynamic action of the Sec machinery during initiation, protein translocation and termination revealed by single molecule fluorescence
Dynamic action of the Sec machinery during initiation, protein translocation and termination revealed by single molecule fluorescence Open
Protein translocation across cell membranes is a ubiquitous process required for protein secretion and membrane protein insertion. This is mediated, for the majority of proteins, by the highly conserved Sec machinery. The bacterial translo…
View article: Targeting of IL-2 to cytotoxic lymphocytes as an improved method of cytokine-driven immunotherapy
Targeting of IL-2 to cytotoxic lymphocytes as an improved method of cytokine-driven immunotherapy Open
The use of high-dose interleukin-2 (IL-2) has fallen out of favor due to severe life-threatening side effects. We have recently described a unique way of directly targeting IL-2 to cytotoxic lymphocytes using a virally encoded immune evasi…
View article: Effect of Bioconjugation on the Reduction Potential of Heme Proteins
Effect of Bioconjugation on the Reduction Potential of Heme Proteins Open
The modification of protein surfaces employing cationic and anionic species enables the assembly of these biomaterials into highly sophisticated hierarchical structures. Such modifications can allow bioconjugates to retain or amplify their…