九州大学 · 材料科学
Hisao Matsuno教授の研究室は、バイオマテリアルと生体分子の相互作用を高感度な質量測定技術(QCM)を用いて解明する研究を展開しています。特に、DNAポリメラーゼやヘリカーゼ、DNaseといった酵素の触媒反応メカニズムをリアルタイムでモニタリングすることで、核酸反応の動的挙動を分子レベルで解明しています。また、ペプチドと高分子の特異的相互作用や、生体適合性ポリマーの界面挙動についても、表面スペクトロスコピーや界面分析を組み合わせた多角的アプローチをとっています。
Figures are computed from collected data and may differ slightly.
Catalytic reactions of DNA polymerase I from E. coli (Klenow fragment, KF) were monitored directly with a template/primer (40/25- or 75/25-mer)-immobilized 27-MHz quartz-crystal microbalance (QCM). The 27-MHz QCM is a very sensitive mass-measuring device in aqueous solution, as the frequency decreases linearly with increasing mass on the QCM electrode at the nanogram level. Three steps in polymerase reactions which include 1) binding of DNA polymerase to the primer on the QCM (mass increase); 2)
Short peptides that recognize the alpha form of poly( l-lactide) (PLLA) crystalline films were identified from a phage-displayed peptide library. An enzyme-linked immunosorbent assay (ELISA) revealed that the apparent binding constants of the phage clones for the alpha form of PLLA were greater than those of the unselected phage library. The specificity index for the alpha form of PLLA referred to a structurally similar atactic poly(methyl methacrylate) (at-PMMA), supporting the alpha form of PL
We have systematically designed and synthesized six kinds of 16-17 mer alanine-based peptides containing four to six lysine (K) and one to four asparagine (N) residues to achieve the selective binding to A.T base pairs of DNA duplexes. The position and number of K and N residues were changed in the helical structure according to common features of the DNA-binding proteins, in which K and N residues are expected to interact electrostatically with phosphate groups and to interact with A.T base pai
In this study, we found that the surface made of a mixture of poly(2-methoxyethyl acrylate) (PMEA) and poly(methyl methacrylate) (PMMA) exhibited excellent blood compatibility by inhibiting platelet adhesion. To obtain a better understanding of this bioinertness, the polymer/water interface was characterized by neutron reflectivity measurements and sum frequency generation spectroscopy, in conjunction with bubble contact angle measurements. Based on the results, we can say that the outermost reg
Catalytic DNA cleavage reactions by an ATP-dependent deoxyribonuclease (DNase) from Micrococcus luteus were monitored directly with a DNA-immobilized 27-MHz quartz-crystal microbalance (QCM). The 27-MHz QCM is a very sensitive mass-measuring device in aqueous solution, as the frequency decreases linearly with increasing mass on the electrode at a nanogram level. Three steps in ATP-dependent DNA hydrolysis reactions, including (1) binding of DNase to the end of double-stranded DNA (dsDNA) on the
Ultrathin poly(methyl methacrylate) (PMMA) stereocomplex films with macromolecularly double-stranded regular nanostructures were prepared by layer-by-layer (LbL) assembly of isotactic (it) and syndiotactic (st) PMMAs on solid substrates, and these films were used for enzyme immobilization supports. Hydrolysis of p-nitrophenyl-β-d-galactopyranoside (PNPG) by the immobilized β-galactosidase (β-Gal) on the complex film was 2- and 4-fold faster than those by the enzyme immobilized on single-componen
We report quantitative analysis of peptide-peptide interactions on a 27 MHz quartz crystal microbalance (QCM) in aqueous solution. The KID (kinase-inducible domain) of transcription factor CREB (cyclic AMP response element binding protein) is known to interact with the KIX domain of coactivator CBP (CREB binding protein), facilitated by phosphorylation at Ser-133 of the KID. The KIX domain peptide (86 aa) was immobilized on the QCM gold electrode surface by means of a poly(ethylene glycol) space
The chain dynamics of well-defined poly(2-methoxyethyl acrylate) (PMEA), which has been used in practice as a bioinert coating for heart-lung machines, was examined as a function of water content by dielectric relaxation spectroscopy (DRS). Two relaxation processes observed in both dried and hydrated films were assigned to the segmental motion (α-process) and the relatively smaller scale motion such as the hindered rotation of side chains (β-process). Water molecules adsorbed on PMEA made the α-
Natural systems, composite materials, and thin-film devices adsorb macromolecules in different phases onto their surfaces. In general, polymer chains form interfacial layers where their aggregation states and thermal molecular motions differ from the bulk. Here, we visualize well-defined double-stranded DNAs (dsDNAs) using atomic force microscopy and molecular dynamics simulations to clarify the adsorption mechanism of polymer chains onto solid surfaces. Initially, short and long dsDNAs are indi
We propose a novel method to treat polymeric scaffold surfaces for cell culture with water containing nanobubbles, called ultrafine bubbles (UFBs), with typical diameters less than 1 μm. A thin film of polystyrene (PS) prepared on a solid substrate was exposed to UFB water for 2 days at room temperature. The PS surface was characterized by X-ray photoelectron spectroscopy (XPS), static contact angle measurements in water, and atomic force microscopy (AFM). The surface chemical composition and we
A star-like hyperbranched polymer having hydrophilic poly(ethyleneoxide acrylate) arms (HB-PEO9A) was prepared by a core-first method based on atom transfer radical polymerization. The PEO9A layer coated on a solid substrate was dissolved by water, and effectively inhibited protein adsorption and cell adhesion.
Solid films of deoxyribonucleic acid (DNA) from salmon testes were prepared by a solvent-casting method. The thermal molecular motion of the DNA film was examined by dynamic mechanical analysis (DMA). Four absorption peaks and one shoulder of the loss modulus were observed in the temperature domain from approximately 150 to 490 K. To assign these, thermal analysis employing thermogravimetry (TG) and differential thermal analysis (DTA) was used in conjunction with ultraviolet (UV)-visible and Fou
Microplastics have recently been identified as one of the major contributors to environmental pollution. To design and control the biodegradability of polymer materials, it is crucial to obtain a better understanding of the aggregation states and thermal molecular motion of polymer chains in aqueous environments. Here, we focus on melt-spun microfibers of a promising biodegradable plastic, polyamide 4 (PA4), with a relatively greater number density of hydrolyzable amide groups, which is regarded
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