Biomedical subjects

Yoshihisa Kaizuka

Publications and source records attributed to Yoshihisa Kaizuka.

4 recordsLinked to original sources

PubMed2006

Hydrodynamic damping of membrane thermal fluctuations near surfaces imaged by fluorescence interference microscopy.

Hydrodynamic coupling between closely apposed lipid bilayer membranes can substantially slow the time scale of thermal fluctuations. These effects could dominate the kinetics of macromolecular interactions at membrane interfaces, and may exert important influences over the interactions between living cells. Here, we study a reconstituted membrane system consisting of two planar bilayers with average separation distances of 30-60 nm. This spacing range is comparable to that of natural intercellular junctions. Using real-time fluorescence interference contrast imaging, spatial and temporal characteristics of the thermal fluctuation spectra are monitored simultaneously. Strong hydrodynamic damping of the fluctuation time scale is observed whereas fluctuation spatial dimensions are unaltered by proximity to another surface. These observations provide experimental confirmation of recent theoretical models.

Lipid Bilayers↗

PubMed2005

Synthesis of lipidated green fluorescent protein and its incorporation in supported lipid bilayers.

Herein we report a semisynthetic method of producing membrane-anchored proteins. Ligation of synthetic lipids with designed anchor structures to proteins was performed using native chemical ligation (NCL) of a C-terminal peptide thioester and an N-terminal cysteine lipid. This strategy mimics the natural glycosylphosphatidylinositol (GPI) linkage found in many natural membrane-associated proteins; however, the synthetic method utilizes simple lipid anchors without glycans. Synthetically lipidated recombinant green fluorescent protein (GFP) was shown to be stably anchored to the membrane, and its lateral fluidity was quantitatively characterized by direct fluorescence imaging in supported membranes. Circumventing the steps of purification from native cell membranes, this methodology facilitates the reconstitution of membrane-associated proteins.

Green Fluorescent Proteins↗

PubMed2005

Formation and spatio-temporal evolution of periodic structures in lipid bilayers.

Miscibility phase separation in lipid bilayers is widely implicated as an organizing principle in living cell membranes. However, the chemical and physical aspects of how membrane phase separation modulates protein activity remain obscure. Herein, we describe formation of ordered superstructures of coexisting liquid phases in bilayer membranes. Metastable stripe and hexagonal domain lattices are observed, as well as transitions between them. The high degree of order achieved by these methods facilitates statistical analysis of domain spatial distributions and enables measurement of domain interactions. Such long-range ordering principles may exist in more complicated membrane systems.

Biomimetic Materials↗

PubMed2004

Structure and dynamics of supported intermembrane junctions.

Supported intermembrane junctions, formed by rupture of giant unilamellar vesicles onto conventional supported lipid membranes, have recently emerged as model systems for the study of biochemical processes at membrane interfaces. Using intermembrane fluorescence resonance energy transfer and optical standing wave fluorescence interferometry, we characterize the nanometer-scale topography of supported intermembrane junctions and find two distinct association states. In one state, the two membranes adhere in close apposition, with intermembrane separations of a few nanometers. In the second state, large intermembrane spacings of approximately 50 nm are maintained by a balance between Helfrich (entropic) repulsion and occasional sites of tight adhesion that pin the two membranes together. Reversible transitions between these two states can be triggered with temperature changes. We further examine the physical properties of membranes in each state using a membrane mixture near its miscibility phase transition temperature. Thermodynamic characteristics of the phase transition and diffusive mobility of individual lipids are comparable. However, collective Brownian motion of phase-separated domains and compositional fluctuations are substantially modulated by intermembrane spacing. The scaling properties of diffusion coefficient with particle size are determined from detailed analysis of domain motion in the different junction types. The results provide experimental verification of a theoretical model for two-dimensional mobility in membranes, which includes frictional coupling across an interstitial water layer.

Computer Simulation↗