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Hayamizu, Y.

Publications and source records attributed to Hayamizu, Y..

2 recordsLinked to original sources

Synthetic spider silk forming highly-aligned nanoarchitectures on 2D-material surfaces

The synergy between silk proteins and nanomaterials can lead to novel materials with improved mechanical and electrical properties. Designed peptides have been previously utilised in the functionalisation of two-dimensional material surfaces in a self-assembly manner, including graphite, to develop highly sensitive electrical biosensors. These studies have predominantly focused on functionalising the surfaces with peptides of less than several kDa in size. In this work, we assessed the capability of a [~]35 kDa synthetic spider silk protein to serve as a candidate biomolecular scaffold on a range of two-dimensional materials: graphite, molybdenum disulphide and boron nitride. The structural properties of the synthetic spider silk protein at the 2D material surface were characterised at the nanoscale for the first time using a multi-analysis approach incorporating atomic force microscopy and fluorescence microscopy, in addition to polarised Raman and tip-enhanced Raman spectroscopy. The synthetic spider silk protein was revealed to self-assemble into stable nanowire structures of monolayer thickness. Our findings have demonstrated the feasibility of functionalising 2D materials with spider silk-based proteins and will unlock new possibilities in the development of next-generation high-performance biosensing devices.

biophysics↗

An improved molecular crowding sensor CRONOS for detection of crowding changes in membrane-less organelles under pathological conditions.

Membrane-less organelles (MLOs) formed by liquid-liquid phase separation (LLPS) play pivotal roles in biological processes. During LLPS, proteins and nucleotides are extremely condensed, resulting in changes of their conformation and biological functions. Disturbed LLPS homeostasis in MLOs cause fatal diseases such as amyotrophic lateral sclerosis. Therefore, it is important to detect changes of the degree of crowding in MLOs. However, it has not been investigated well due to lack of an appropriate method. To address this, we developed a genetically-encoded molecular crowding sensor CRONOS that senses the degree of macromolecular crowding in MLOs using fluorescence resonance energy transfer (FRET) system. CRONOS is a very bright biosensor with wider dynamic range and detect changes in the macromolecular volume fraction better than the previously reported mCer-mCit sensor in solution. By fusing to scaffold protein of each MLO, we successfully delivered CRONOS to MLO of interest and detected previously undescribed difference of the degree of crowding in each MLO. If not tagged, CRONOS localized to interstitial space of MLOs, giving us the crowding information of inspace. CRONOS also detected changes of degree of macromolecular crowding in nucleolus induced by environmental stress or inhibition of transcription. These findings suggest that CRONOS can be a useful tool for determination of molecular crowding and detection of pathological changes in MLOs in live cells.

biochemistry↗