A5044554319- Cellular Mechanics and Interactions
- Monoclonal and Polyclonal Antibodies Research
- Biochemical and Structural Characterization
- SARS-CoV-2 and COVID-19 Research
- SARS-CoV-2 detection and testing
- Cell Adhesion Molecules Research
- 3D Printing in Biomedical Research
- COVID-19 Clinical Research Studies
- Bone Tissue Engineering Materials
- RNA and protein synthesis mechanisms
- Electrospun Nanofibers in Biomedical Applications
- Force Microscopy Techniques and Applications
- vaccines and immunoinformatics approaches
- Bacterial Infections and Vaccines
- Transgenic Plants and Applications
- Nanofabrication and Lithography Techniques
- Protease and Inhibitor Mechanisms
- Light effects on plants
- Biotin and Related Studies
- Heat shock proteins research
- Glycosylation and Glycoproteins Research
- Polymer Surface Interaction Studies
- Histone Deacetylase Inhibitors Research
- Viral gastroenteritis research and epidemiology
- Cellular transport and secretion
Tampere University
2014-2024
Fimlab (Finland)
2014-2023
University of Oxford
2019-2022
Pirkanmaa Hospital District
2014
Much of life's complexity depends upon contacts between proteins with precise affinity and specificity. The successful application engineered often on high-stability binding to their target. In recent years, various approaches have enabled form irreversible covalent interactions protein targets. However, the rate such reactions is a major limitation use. Infinite refers ideal where interaction occurs at diffusion limit. Prototypes infinite pairs been achieved using nonnatural reactive...
Abstract There is need for effective and affordable vaccines against SARS-CoV-2 to tackle the ongoing pandemic. In this study, we describe a protein nanoparticle vaccine SARS-CoV-2. The based on display of coronavirus spike glycoprotein receptor-binding domain (RBD) synthetic virus-like particle (VLP) platform, SpyCatcher003-mi3, using SpyTag/SpyCatcher technology. Low doses RBD-SpyVLP in prime-boost regimen induce strong neutralising antibody response mice pigs that superior convalescent...
Abstract Matching of symmetry at interfaces is a fundamental obstacle in molecular assembly. Virus‐like particles (VLPs) are important vaccine platforms against pathogenic threats, including Covid‐19. However, mismatch can prohibit nanoassembly. We established an approach for coupling VLPs to diverse antigen symmetries. SpyCatcher003 enabled efficient VLP conjugation and extreme thermal resilience. Many people had pre‐existing antibodies SpyTag:SpyCatcher but less the 003 variants. coupled...
Abstract Serological detection of antibodies to SARS-CoV-2 is essential for establishing rates seroconversion in populations, and seeking evidence a level antibody that may be protective against COVID-19 disease. Several high-performance commercial tests have been described, but these require centralised laboratory facilities are comparatively expensive, therefore not available universally. Red cell agglutination do special equipment, read by eye, short development times, low cost can...
Abstract Poly(vinylidene fluoride) (PVDF) is a biocompatible material with excellent electroactive properties. Nonelectroactive α‐PVDF and β‐PVDF were used to investigate the substrate polarization polarity influence on focal adhesion (FA) size number as well human adipose stem cells (hASCs) differentiation. hASCs cultured different PVDF surfaces adsorbed fibronectin FA number, total area, cell size, aspect ratio density estimated using expressing vinculin fused enhanced green fluorescent...
The mechanical unfolding of proteins is a cellular mechanism for force transduction with potentially broad implications in cell fate. Despite this, the by which protein elicits differential downstream signalling pathways remains poorly understood. Here, we used engineering, atomic microscopy, and biophysical tools to delineate how controls mechanics. Deleted liver cancer 1 (DLC1) negative regulator Ras homolog family member A (RhoA) contractility that regulates behaviour when localised focal...
Talin protein is one of the key components in integrin-mediated adhesion complexes. Talins transmit mechanical forces between β-integrin and actin, regulate complex composition signaling through force-regulated unfolding talin rod domain. Using modified proteins, we demonstrate that these functions contribute to different cellular processes can be dissected. The transmission regulates phosphotyrosine even absence mechanically regulated subdomains. However, presence subdomains their...
Cells adhere to the surrounding tissue and probe its mechanical properties by forming cell-matrix adhesions. Talin is a critical adhesion protein participates in transmission of signals between extracellular matrix cell cytoskeleton. Force induced unfolding talin rod subdomains has been proposed act as cellular mechanosensor, but so far evidence linking their stability response lacking. Here, utilizing computationally designed mutations, we demonstrate that stepwise destabilization R3...
Integrin-dependent cell adhesion and spreading are critical for morphogenesis, tissue regeneration, immune defense but also tumor growth. However, the mechanisms that induce integrin-mediated provide mechanosensing on different extracellular matrix conditions not fully understood. By expressing β3-GFP-integrins with enhanced talin-binding affinity, we experimentally uncoupled integrin activation, clustering, substrate binding from its function in spreading. Mutational analysis revealed...
ABSTRACT There is dire need for an effective and affordable vaccine against SARS-CoV-2 to tackle the ongoing pandemic. In this study, we describe a modular virus-like particle candidate displaying spike glycoprotein receptor-binding domain (RBD) using SpyTag/SpyCatcher technology (RBD-SpyVLP). Low doses of RBD-SpyVLP in prime-boost regimen induced strong neutralising antibody response mice pigs that was superior convalescent human sera. We evaluated quality ACE2 blocking neutralisation cell...
Abstract Proteins can be empowered via SpyTag for anchoring and nanoassembly, through covalent bonding to SpyCatcher partners. Here we generate a switchable version of SpyCatcher, allowing gentle purification SpyTagged proteins. We introduce numerous histidines adjacent SpyTag’s binding site, giving moderate pH-dependent release. After phage-based selection, our final SpySwitch allows SpyTag- SpyTag003-fusions from bacterial or mammalian culture by capture at neutral pH release 5, with...
ABSTRACT Integrin activation and clustering by talin are early steps of cell adhesion. Membrane-bound head domain kindlin bind to the β integrin cytoplasmic tail, cooperating activate heterodimeric integrin, induces in presence Mn2+. Here we show that kindlin-1 can replace Mn2+ mediate β3 induced head, but not F2–F3 fragment talin. mediated was lost upon deletion flexible loop within F1 subdomain. Further mutagenesis identified hydrophobic acidic motifs responsible for clustering. Modeling,...
Talin (herein referring to the talin-1 form), is a cytoskeletal adapter protein that binds integrin receptors and F-actin, key factor in formation regulation of integrin-dependent cell-matrix adhesions. forms mechanical link between cytoplasmic domain integrins actin cytoskeleton. Through this linkage, talin at origin mechanosignaling occurring plasma membrane-cytoskeleton interface. Despite its central position, not able fulfill tasks alone, but requires help from kindlin paxillin detect...
The traditional silicate bioactive glasses exhibit poor thermal processability, which inhibits fiber drawing or sintering into scaffolds. composition of the has been modified to enable hot processing. However, forming ability is generally at expense bioactivity. Metaphosphate glasses, on other hand, possess excellent congruent dissolution, and a tailorable degradation rate. due layer-by-layer dissolution mechanism, cells do not attach material surface. Furthermore, leads low density OH...
Light is well-established for control of bond breakage but not specific formation in complex environments. We previously engineered the diffusion-limited reactivity SpyTag003 peptide with its protein partner SpyCatcher003 through spontaneous isopeptide formation. This system enables precise and irreversible assembly biological building blocks applications from biomaterials to vaccines. Here we establish a rapid this amide visible light. have generated caged SpyCatcher003, which allows light...
The application of nanotechnology in biomedical field has enormous potential basic and applied research. Micro or nanofibers produced by electrospinning technique offer excellent properties because large specific surface area, high porosity, ability to incorporate functional additives. Here we embedded biotinylated bovine serum albumin into polylactic acid (PLA)-polyethylene glycol (PEG) fibers, which enabled immobilization fluorescently labelled avidin. An alkaline phosphatase enzyme was...
Matching of symmetry at interfaces is a fundamental obstacle in molecular assembly. Virus-like particles (VLPs) are important vaccine platforms against pathogenic threats, including Covid-19. However, mismatch can prohibit nanoassembly. We established an approach for coupling VLPs to diverse antigen symmetries. SpyCatcher003 enabled efficient VLP conjugation and extreme thermal resilience. Many people had pre-existing antibodies SpyTag:SpyCatcher but less the 003 variants. coupled...
Vinculin binds to specific sites of mechanically unfolded talin rod domains reinforce the coupling cell’s exterior its force generation machinery. Force-dependent vinculin–talin complexation and dissociation was previously observed as contraction or extension respectively using magnetic tweezers. However, structural mechanism underlying vinculin recognition binding (VBSs) in remains unknown. Using molecular dynamics simulations, we demonstrate that a VBS dynamically refolds under force, can...
Avidins are a family of proteins widely employed in biotechnology. We have previously shown that functional chimeric mutant can be created from avidin and avidin-related protein 2 using methodology combining random mutagenesis by recombination selection tailored biopanning protocol (phage display). Here, we report the crystal structure one selected characterized forms, A/A2-1. The was solved at 1.8 Å resolution revealed fold not affected shuffled sequences. also supports observed...
ABSTRACT Serological detection of antibodies to SARS-CoV-2 is essential for establishing rates seroconversion in populations, after vaccination, and seeking evidence a level antibody that may be protective against COVID-19 disease. Several high-performance commercial tests have been described, but these require centralised laboratory facilities are comparatively expensive, therefore not available universally. Red cell agglutination long history blood typing, general serology through linkage...
Efficient and robust subcloning is essential for the construction of high-diversity DNA libraries in field directed evolution. We have developed a more efficient method DNA-shuffled by employing recombination cloning (Gateway). The Gateway procedure was performed directly after gene reassembly reaction, without additional purification amplification steps, thus simplifying conventional shuffling protocols. Recombination-based cloning, from heterologous conserved high quality library reduced...
Abstract Serological detection of antibodies to SARS-CoV-2 is essential for establishing rates seroconversion in populations, after vaccination, and seeking evidence a level antibody that may be protective against COVID-19 disease. Several high-performance commercial tests have been described, but these require centralised laboratory facilities are comparatively expensive, therefore not available universally. Red cell agglutination long history blood typing, general serology through linkage...