A5004297533- Endoplasmic Reticulum Stress and Disease
- Viral Infectious Diseases and Gene Expression in Insects
- Protein Structure and Dynamics
- Receptor Mechanisms and Signaling
- Advanced biosensing and bioanalysis techniques
- Insect symbiosis and bacterial influences
- CRISPR and Genetic Engineering
- RNA and protein synthesis mechanisms
- Enzyme Structure and Function
- Bacterial Genetics and Biotechnology
- Insect and Pesticide Research
- Biotin and Related Studies
- Mosquito-borne diseases and control
- Electrochemical sensors and biosensors
- Cytokine Signaling Pathways and Interactions
- Monoclonal and Polyclonal Antibodies Research
- Molecular Junctions and Nanostructures
- Adenosine and Purinergic Signaling
- Microbial Fuel Cells and Bioremediation
- Anodic Oxide Films and Nanostructures
- Cellular transport and secretion
- Myeloproliferative Neoplasms: Diagnosis and Treatment
- Calcium Carbonate Crystallization and Inhibition
- Pharmacological Effects of Natural Compounds
- Protein purification and stability
Queensland University of Technology
2019-2025
Technology Centre Prague
2025
ARC Centre of Excellence in Synthetic Biology
2019-2023
Commonwealth Scientific and Industrial Research Organisation
2019-2023
The University of Queensland
2011-2019
Monash University
2009
Allosteric protein switches are key controllers of information and energy processing in living organisms desirable engineered control tools synthetic systems. Here we present a generally applicable strategy for construction allosteric signaling systems with inputs outputs choice. We demonstrate conversion constitutively active enzymes into peptide-operated ON by insertion calmodulin domain rationally selected sites. Switches based on EGFP, glucose dehydrogenase, NanoLuciferase, dehydrofolate...
Allostery enables proteins to interconvert different biochemical signals and form complex metabolic signaling networks. We hypothesize that circular permutation of increases the probability functional coupling new N- C- termini with protein's active center through increased local structural disorder. To test this we construct a synthetically allosteric version permutated NanoLuc luciferase can be activated ligand-induced intramolecular non-covalent cyclisation. This switch module is tolerant...
Lanthanides, which are part of the rare earth elements group have numerous applications in electronics, medicine and energy storage. However, our ability to extract them is not meeting rapidly increasing demand. The discovery bacterial periplasmic lanthanide‐binding protein lanmodulin spurred significant interest developing biotechnological routes for lanthanide detection extraction. Here we report construction b‐lactamase‐lanmodulin chimeras that function as lanthanide‐controlled enzymatic...
Lanthanides, which are part of the rare earth elements group have numerous applications in electronics, medicine and energy storage. However, our ability to extract them is not meeting rapidly increasing demand. The discovery bacterial periplasmic lanthanide‐binding protein lanmodulin spurred significant interest developing biotechnological routes for lanthanide detection extraction. Here we report construction b‐lactamase‐lanmodulin chimeras that function as lanthanide‐controlled enzymatic...
Abstract Protein biosensors play an increasingly important role as reporters for research and clinical applications. Here we present approach the construction of fully integrated but modular electrochemical based on principal component glucose monitors PQQ‐glucose dehydrogenase (PQQ‐GDH). We designed allosterically regulated circular permutated variants PQQ‐GDH that show large (>10‐fold) changes in enzymatic activity following intramolecular scaffolding newly generated N‐ C termini by...
Abstract Natural evolution produced polypeptides that selectively recognize chemical entities and their polymers, ranging from ions to proteins nucleic acids. Such selective interactions serve as entry points biological signaling metabolic pathways. The ability engineer artificial versions of such is a key goal synthetic biology, bioengineering bioelectronics. We set out map the optimal strategy for developing small molecule:protein complexes function chemically induced dimerization (CID)...
The bacterial disulfide machinery is an attractive molecular target for developing new antibacterials because it required the production of multiple virulence factors. archetypal oxidase proteins in Escherichia coli (Ec) are DsbA and DsbB, which together form a functional unit: introduces disulfides into folding DsbB reoxidizes to maintain active form. In Mycobacterium tuberculosis (Mtb), no homologue encoded but functionally similar structurally divergent protein, MtbVKOR, has been...
The ability of proteins to interconvert unrelated biochemical inputs and outputs underlays most energy information processing in biology. A common conversion mechanism involves a conformational change protein receptor response ligand binding or covalent modification, leading allosteric activity modulation the effector domain. Designing such systems rationally is central goal synthetic biology engineering. two-component sensory system based on scaffolding modules presence an analyte one...
The enzyme TcpG is a periplasmic protein produced by the Gram-negative pathogen Vibrio cholerae. essential for production of ToxR-regulated proteins, including virulence-factor pilus proteins and cholera toxin, therefore target development new class anti-virulence drugs. Here, 1.2 Å resolution crystal structure reported using cryocooled crystal. This compared with previous determined at 2.1 from data measured room temperature. first DsbA to be solved sufficiently high allow inclusion refined...
The emergence of viral threats such as Ebola, ZIKA, and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) requires a rapid efficient approach for elucidating mechanisms pathogenesis development therapeutics. In this context, cell-free protein synthesis (CFPS) holds promise to resolve the bottlenecks multiplexed production interaction analysis among host pathogen proteins. Here, we applied eukaryotic CFPS system based on Leishmania tarentolae extract (LTE) expression in combination...
The rapid spread of arthropod-borne Zika virus poses a serious public health threat that calls for effective ways controlling and treating viral infection. This in turn necessitates better understanding the mechanisms assembly its interaction with host cells. In order to facilitate such efforts, we developed new multihost expression vector pmCellFree allows multiplexed production ZIKV proteins any vitro translation system as well mammalian Using combination Leishmania cell-free AlphaLISA...
Despite the significant potential of protein biosensors, their construction remains a trial-and-error process. The most obvious approach for addressing this is to utilize modular biosensor architectures where specificity-conferring modalities can be readily generated recognize new targets. Toward goal, we established workflow that uses mRNA display-based selection hyper-stable monobody domains target choice or ribosome display select equally stable DARPins. These binders were integrated into...
The α-proteobacterium Wolbachia pipientis infects more than 65% of insect species worldwide and manipulates the host reproductive machinery to enable its own survival. It can live in mutualistic relationships with hosts that cause human disease, including mosquitoes carry Dengue virus. Like many other bacteria, contains disulfide bond forming (Dsb) proteins introduce bonds into secreted effector proteins. genome strain wMel encodes two DsbA-like sharing just 21% sequence identity each other,...
Enzymatic polypeptide proteolysis is a widespread and powerful biological control mechanism. Over the last few years, substantial progress has been made in creating artificial proteolytic systems where an input of choice modulates protease activity thereby its substrates. However, all developed so far have relied on direct cleavage their effectors. Here, we propose new concept biosensors with tunable uncage signaling peptide, which can then transmit signal to allosteric protein reporter. We...
Disulfide-bond-forming (DSB) oxidative folding enzymes are master regulators of virulence that localized to the periplasm many Gram-negative bacteria. The archetypal DSB machinery from Escherichia coli K-12 consists a dithiol-oxidizing redox-relay pair (DsbA/B), disulfide-isomerizing (DsbC/D) and specialist reducing DsbE DsbG also interact with DsbD. By contrast, bacterium Wolbachia pipientis encodes just three enzymes. Two these, α-DsbA1 α-DsbB, form analogous DsbA/B E. . third enzyme,...
Antibiotic resistance is a major global health threat. Bacteria have developed novel mechanisms to many of the latest generations antibiotics and there an urgent need develop new therapies combat these infections. Infections that are caused by multi-drug resistant Gram-negative bacteria result in poor prognosis, prolonged illness, greater costs for care. Recent research has pointed several key bacterial membrane proteins as potential targets drug vaccine development. However, determination...
Abstract Protein biosensors play an increasingly important role as reporters for research and clinical applications. Here we present approach the construction of fully integrated but modular electrochemical based on principal component glucose monitors PQQ‐glucose dehydrogenase (PQQ‐GDH). We designed allosterically regulated circular permutated variants PQQ‐GDH that show large (>10‐fold) changes in enzymatic activity following intramolecular scaffolding newly generated N‐ C termini by...