A5034545835- Enzyme Structure and Function
- RNA and protein synthesis mechanisms
- Protein Structure and Dynamics
- Advancements in Battery Materials
- Polyoxometalates: Synthesis and Applications
- Luminescence Properties of Advanced Materials
- Enzyme Production and Characterization
- Nanocluster Synthesis and Applications
- Hemoglobin structure and function
- Protein Kinase Regulation and GTPase Signaling
- Glycosylation and Glycoproteins Research
- Chemical Synthesis and Analysis
- Advanced Battery Materials and Technologies
- Galectins and Cancer Biology
- Plant Genetic and Mutation Studies
- Analytical Chemistry and Sensors
- Genomics and Phylogenetic Studies
- RNA modifications and cancer
- Advanced Battery Technologies Research
- DNA Repair Mechanisms
- Carcinogens and Genotoxicity Assessment
- Metalloenzymes and iron-sulfur proteins
- Peptidase Inhibition and Analysis
- Microwave Dielectric Ceramics Synthesis
- Bacteriophages and microbial interactions
KU Leuven
2017-2022
Yokohama City University
2012-2021
Saitama University
2018
Indiana University School of Medicine
2017
Indiana University – Purdue University Indianapolis
2017
Saga University
1978-2009
Aichi Medical University
2009
Okayama University of Science
1986
Significance In this study, we have designed and experimentally validated, to our knowledge, the first perfectly symmetrical β-propeller protein. Our results provide insight not only into protein evolution through duplication events, but also methods for creating designer proteins that self-assemble according simple arithmetical rules. Such may very wide uses in bionanotechnology. Furthermore design approach is both rapid applicable many different templates. novel propeller consists of six...
Unraveling the mechanism of action and molecular target small molecules remains a major challenge in drug discovery. While many cancer drugs genetic vulnerabilities, loss-of-function screens fail to identify essential genes action. Here, we report CRISPRres, CRISPR-Cas-based screening approach rapidly derive resistance mutations genes. It exploits local variation created by CRISPR-Cas-induced non-homologous end-joining (NHEJ) repair generate wide variety functional in-frame mutations. Using...
The five human RecQ helicases participate in multiple processes required to maintain genome integrity. Of these, the disease-linked RecQ4 is least studied because it poses many technical challenges. We previously demonstrated that yeast Hrq1 helicase displays similar functions vivo, and here, we report biochemical structural characterization of these enzymes. In vitro, are DNA-stimulated ATPases robust helicases. Further, activities were sensitive DNA sequence structure, with preferentially...
MytiLec is a lectin, isolated from bivalves, with cytotoxic activity against cancer cell lines that express globotriaosyl ceramide, Galα(1,4)Galβ(1,4)Glcα1-Cer, on the surface. Functional analysis shows protein binds to disaccharide melibiose, Galα(1,6)Glc, and trisaccharide globotriose, Galα(1,4)Galβ(1,4)Glc. Recombinant expressed in bacteria showed same haemagglutinating Burkitt's lymphoma (Raji) cells as native form. The crystal structure has been determined atomic resolution, presence...
Abstract Computational protein design has advanced very rapidly over the last decade, but there remain few examples of artificial proteins with direct medical applications. This study describes a new β -trefoil lectin that recognises Burkitt’s lymphoma cells, and which was designed intention finding basis for novel cancer treatments or diagnostics. The protein, called “Mitsuba”, is based on structure natural shellfish MytiLec-1, member small family uses unique sequence motifs to bind α...
Abstract We have engineered a metal‐binding site into the novel artificial β‐propeller protein Pizza. This new Pizza variant carries two nearly identical domains per polypeptide chain, and forms trimer with three‐fold symmetry. The designed single metal ion binding lies on symmetry axis, bonding together. Two copies of associate in presence cadmium chloride solution, very high‐resolution X‐ray crystallographic analysis reveals nanocrystal chloride, sandwiched between trimers protein....
β-Propeller proteins form one of the largest families protein structures, with a pseudo-symmetrical fold made up subdomains called blades. They are not only abundant but also involved in wide variety cellular processes, often by acting as platform for assembly complexes. WD40 subfamily propeller no intrinsic enzymatic activity, their stable, modular architecture and versatile surface have allowed evolution to adapt them many vital roles. By computationally reverse-engineering duplication,...
A symmetric designer protein forms hybrid complexes with different polyoxometalates and may serve as a building block for porous frameworks.
The controlled formation of protein supramolecular assemblies is challenging, but it could provide an important route for the development hybrid biomaterials. In this work, we demonstrate well-defined complexes between eightfold symmetrical designer Tako8 and soluble metal-oxo clusters from family Anderson–Evans, Keggin, ZrIV-substituted Wells–Dawson polyoxometalates. A combination X-ray crystallography solution studies showed that are able to serve as linker nodes bottom-up creation...
Bacterial biofilm formation is an extremely widespread phenomenon involving the secretion of a protective exopolysaccharide matrix which helps bacteria to attach surfaces and overcome variety stresses in different environments. This may also include proteins, lipids, DNA metal ions. Its composition depends on bacterial species growth conditions, but one most widely found components polymeric β-1,6-N-acetyl-D-glucosamine (PGA). Several studies have suggested that PGA essential component it...
β‐propeller proteins are common in nature, where they observed to adopt 4‐ 10‐fold internal rotational pseudo‐symmetry. This size diversity can be explained by the evolutionary process of gene duplication and fusion. In this study, we investigated a distorted protein, an apparent intermediate between two symmetries. From template, created perfectly symmetric 9‐bladed named Cake, using computational design ancestral sequence reconstruction. The designed repeat was found capable generating...
The β-propeller fold is adopted by a sequentially diverse family of repeat proteins with apparent rotational symmetry. While the structure mostly stabilized hydrophobic interactions, an additional stabilization provided hydrogen bonds between N-and C-termini, which are almost invariably part same β-sheet. This feature often referred to as "Velcro" closure. positioning termini within blade variable and depends on protein family. In order investigate influence this location structure, folding...
Type-2 diabetes is characterized by obesity-related insulin resistance. Insulin resistance and accompanying hyperinsulinemia have been reported to play an important role in pathogenesis of the metabolic syndrome. Conjugated linoleic acid (CLA), a mixture positional geometric isomers acid, has attracted considerable attention because its potentially beneficial biological effects. Previous studies showed that dietary CLA alleviates through improvement glucose tolerance insulin-stimulated...
Rap1B is a small GTPase involved in the regulation of numerous cellular processes including synaptic plasticity, one bases memory. Like other members Ras family, active GTP-bound form can bind to large number effector proteins and so transmit signals downstream components signaling pathways. The structure bound only nucleotide has yet be solved, but might help reveal an inactive conformation that stabilized by molecule drug. Unlike family such as H-Ras Rap2A, crystallizes intermediate state...
We developed an artificial hydrolase based on the symmetrical Pizza6 β-propeller protein for metal-free hydrolysis of 4-nitrophenyl acetate and butyrate. Through site-specific mutagenesis crystallisation studies, catalytic mechanism was investigated found to be dependent a threonine-histidine dyad. The mutant with additional histidine residues generated highest kcat values, forming His-His-Thr triad matched previously reported metalloenzymes. highly enzymes their protein-metal complexes have...
The haemoglobin (Hb) of the extinct woolly mammoth has been recreated using recombinant genes expressed in Escherichia coli. globin gene sequences were previously determined DNA recovered from frozen cadavers. Although highly similar to Hb existing elephants, protein shows rather different responses chloride ions and temperature. In particular, heat oxygenation is found be much lower Hb, which appears an adaptation harsh high-latitude climates Pleistocene Ice Ages linked heightened...
Certain experiments on the destructive action of bile and salts upon pancreas made by Flexner indicated that colloidal substances restrained salts. It was suggested this restraint probably depended a reduction in rapidity diffusion into tissues. The studies Voigtländer influence colloids (agar-agar) rate certain crystalloids tended to show agar-agar jelly takes place at about same as water. summarized communication were with hemolytic suspended isotonic saline solution gelatin jelly. could...
Abstract The effects of the ammonium-crown ether or potassium-crown complex on extraction anionic species were studied. cobalt thiocyanate was extracted into 1,2-dichloroethane by addition a crown complex. By measuring absorbance organic phase at 623 nm, determined spectrophotometrically. A linear calibration curve obtained up to 10−4 M in aqueous phase.
Abstract Crown ether-potassium ion complex extracts selectively copper-zincon anion as ion-pairs into chloroform phase in the presence of and zinc-zincon chelate anion, whereas quarternary ammonium both anions. By measuring absorbance at 615 nm, copper zinc has been determined spectrophotometrically. The composition extracted species was estimated to be Cu2+-zincon4−-(KL+)2.