A5037114896- Enzyme Structure and Function
- Amino Acid Enzymes and Metabolism
- Biochemical and Molecular Research
- Antioxidant Activity and Oxidative Stress
- Retinoids in leukemia and cellular processes
- Polyamine Metabolism and Applications
- Glycosylation and Glycoproteins Research
- Metabolism and Genetic Disorders
- Protein Structure and Dynamics
- Estrogen and related hormone effects
- Enzyme Catalysis and Immobilization
- Microbial Metabolic Engineering and Bioproduction
- Phytochemicals and Antioxidant Activities
- Erythrocyte Function and Pathophysiology
- Enzyme Production and Characterization
- Monoclonal and Polyclonal Antibodies Research
- Tea Polyphenols and Effects
- Diabetes Treatment and Management
- Advanced Glycation End Products research
- Chemical Synthesis and Analysis
- Peroxisome Proliferator-Activated Receptors
- Peptidase Inhibition and Analysis
- Inflammatory mediators and NSAID effects
- X-ray Diffraction in Crystallography
- Protein Interaction Studies and Fluorescence Analysis
University of Shizuoka
2015-2024
Japan Agency for Medical Research and Development
2020
Shizuoka University
2018
Food & Nutrition
2011-2017
National Fisheries University
2014
The University of Tokyo
2001-2013
Nagoya University
2009
University of Milan
2009
Proximity biotinylation based on Escherichia coli BirA enzymes such as BioID (BirA*) and TurboID is a key technology for identifying proteins that interact with target protein in cell or organism. However, there have been some improvements the are used purpose. Here, we demonstrate novel enzyme, AirID (ancestral proximity-dependent biotin identification), which was designed de novo using an ancestral enzyme reconstruction algorithm metagenome data. AirID-fusion AirID-p53 AirID-IκBα indicated...
Human serum albumin (HSA) contributes to the stabilization of (−)-epigallocatechin gallate (EGCg) in serum. We characterize present study mechanisms for preventing EGCg oxidation by HSA. was stable human or buffers with HSA, but (EGC) unstable. show comparing and EGC a neutral buffer that had higher binding affinity than EGC. This indicates galloyl moiety participated interaction HSA this critical importance oxidation. The protein carbonyl formation were enhanced an alkaline buffer. These...
Enantiomerically pure amino acid derivatives could be foundational compounds for peptide drugs. Deracemization of racemates to l-amino can achieved through the reaction evolved d-amino oxidase and chemical reductants, whereas deracemization has not progressed due difficulty associated with heterologous expression (LAAO). In this study, we succeeded in developing an ancestral LAAO (AncLAAO) bearing broad substrate selectivity (13 acids) high productivity Escherichia coli system (∼50.7 mg/L)....
Human serum albumin (HSA), the most abundant protein in plasma, has a very unique function, catalyzing conversion of prostaglandin J2 (PGJ2), dehydration product PGD2, to yield Δ12-PGJ2. These PGD2 metabolites are actively transported into cells and accumulated nuclei, where they act as potent inducers cell growth inhibition differentiation, exhibit their own spectrum biological effects. The facts that (i) arachidonic acid bind human (HSA) metabolism these molecules is altered result...
4-Hydroxy-2-nonenal (HNE), a major racemic product of lipid peroxidation, preferentially reacts with cysteine residues to form stable HNE-cysteine Michael addition adduct possessing three chiral centers. Here, gain more insight into sulfhydryl modification by HNE, we characterized the stereochemical configuration adducts and investigated their stereoselective formation in redox-regulated proteins. To characterize NMR, authentic (R)-HNE- (S)-HNE-cysteine were prepared incubating...
L-amino acid oxidases (LAAOs) can be applied to convert racemic amino acids D-isomers, which are potential precursors of pharmaceuticals. However, this application is hampered by the lack available stable and structure-determined LAAOs. In study, we attempt address limitation utilizing two ancestral LAAOs: AncLAAO-N4 AncLAAO-N5. has highest thermal temporal stabilities among designed LAAOs that used for deracemization stereoinversion. AncLAAO-N5 provide X-ray crystal structures, helpful...
Several enzymes are already used on the industrial scale because of development various enzyme engineering approaches. In this study, we developed a protein redesign tool, GAOptimizer, to tackle challenge. GAOptimizer is genetic algorithm-based tool for optimizing mutation combinations engineer diverse enzymes. The requires two input parameters influencing selection: fitness functions and sequence libraries. Both stability-based non-stability-based scores can serve as functions, these...
The pyruvate kinase (PK) from a moderate thermophile, Geobacillus stearothermophilus, is an allosteric enzyme activated by AMP and ribose 5-phosphate but not fructose 1, 6-bisphosphate (FBP), which common activator of PKs. It has extra C-terminal sequence (ECTS), contains highly conserved phosphoenolpyruvate (PEP) binding motif, its function structure remain unclear. To elucidate the structural characteristics effector-binding site ECTS, crystal C9S/C268S mutant was determined at 2.4 Å...
Quercus species occur widely in East Asia and are major tree Japan. The ground-grown seedlings of serrata mongolica var. crispula were exposed to enriched ozone (ambient + 40 ppbv) over one growing season by using open-top-chambers (OTC), isoprene emissions from the two tree-species measured across a leaf cuvette method. emission rates significantly affected ozone, rate Q. was reduced greater extent October than that serrata. stomatal conductance higher not O3 even September, suggesting...
In this study, we attempted to infer the molecular evolution of a recently isolated FAD-dependent l -arginine oxidase (AROD) that oxidizes 2-ketoarginine. Utilizing 10 candidate AROD sequences, obtained total three ancestral ARODs. addition, one native was by cloning The sequences were selected utilizing curation method defined in study. All ARODs successfully expressed Escherichia coli for analysis their biochemical functions. catalytic activity our bacterially suggests ASR successful. is...
Production of D-amino acids (D-AAs) on a large-scale enables to provide precursors peptide therapeutics. In this study, we designed novel L-amino acid oxidase, HTAncLAAO2, by ancestral sequence reconstruction, exhibiting high thermostability and long-term stability. The crystal structure HTAncLAAO2 was determined at 2.2 Å X-ray crystallography, revealing that the enzyme has an octameric form like "ninja-star" feature. Enzymatic property analysis demonstrated exhibits three-order larger...
Several lines of evidence indicate that the nonenzymatic oxidative modification proteins and subsequent accumulation modified have been found in cells during aging stress various pathological states, including premature diseases, muscular dystrophy, rheumatoid arthritis, atherosclerosis. Our previous work suggested existence molecular mimicry between antibodies raised against hydroxy-2-nonenal (HNE)-modified protein anti-DNA autoantibodies, a serologic hallmark systemic lupus erythematosus...
Abstract We have recently reported that green soybean cultivar, echigomidori , and not the yellow fukuyutaka is a rich source of hormone-like peptide leginsulin consisting 37 amino acids (Leg_1_37, PDB 1JU8A) its C-terminal glycine deletant, Leg_1_36. Green mature, but color seedcoat cotyledon remains green. Therefore, in this study, we examined content different varieties 11 colored soybeans (including green, yellow, red, brown black) edamame (immature soybean). Profile analysis...
The selective PPARα modulator (SPPARMα) is expected to medicate dyslipidemia with minimizing adverse effects. Recently, pemafibrate was screened from the ligand library as an SPPARMα bearing strong potency. Several clinical pieces of evidence have proved usefulness a medication; however, how works at molecular level not fully known. In this study, we investigate mechanism behind its novel character through combination approaches X-ray crystallography, isothermal titration calorimetry (ITC),...