scholarly journals High-Speed AFM Images of Thermal Motion Provide Stiffness Map of Interfacial Membrane Protein Moieties

Nano Letters ◽  
2014 ◽  
Vol 15 (1) ◽  
pp. 759-763 ◽  
Author(s):  
Johannes Preiner ◽  
Andreas Horner ◽  
Andreas Karner ◽  
Nicole Ollinger ◽  
Christine Siligan ◽  
...  
Structure ◽  
2019 ◽  
Vol 27 (1) ◽  
pp. 152-160.e3 ◽  
Author(s):  
Takamitsu Haruyama ◽  
Yasunori Sugano ◽  
Noriyuki Kodera ◽  
Takayuki Uchihashi ◽  
Toshio Ando ◽  
...  

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Takanobu A. Katoh ◽  
Takashi Daiho ◽  
Kazuo Yamasaki ◽  
Stefania Danko ◽  
Shoko Fujimura ◽  
...  

AbstractThe sarcoendoplasmic reticulum Ca2+-ATPase (SERCA) transports Ca2+ ions across the membrane coupled with ATP hydrolysis. Crystal structures of ligand-stabilized molecules indicate that the movement of actuator (A) domain plays a crucial role in Ca2+ translocation. However, the actual structural movements during the transitions between intermediates remain uncertain, in particular, the structure of E2PCa2 has not been solved. Here, the angle of the A-domain was measured by defocused orientation imaging using isotropic total internal reflection fluorescence microscopy. A single SERCA1a molecule, labeled with fluorophore ReAsH on the A-domain in fixed orientation, was embedded in a nanodisc, and stabilized on Ni–NTA glass. Activation with ATP and Ca2+ caused angle changes of the fluorophore and therefore the A-domain, motions lost by inhibitor, thapsigargin. Our high-speed set-up captured the motion during EP isomerization, and suggests that the A-domain rapidly rotates back and forth from an E1PCa2 position to a position close to the E2P state. This is the first report of the detection in the movement of the A-domain as an angle change. Our method provides a powerful tool to investigate the conformational change of a membrane protein in real-time.


2016 ◽  
Author(s):  
David Albrecht ◽  
Christian M. Winterflood ◽  
Thomas Tschager ◽  
Helge Ewers

AbstractThe axon initial segment (AIS) is enriched in specific adaptor, cytoskeletal and transmembrane molecules. During AIS establishment, a membrane diffusion barrier is formed between the axon and the somatodendritic domain. Recently, an axonal periodic pattern of actin, spectrin and ankyrin forming 190 nm distanced, ring-like structures has been discovered. However, whether this structure is related to the diffusion barrier function is unclear.Here, we performed single particle tracking timecourse experiments on hippocampal neurons during AIS development. We analyzed the mobility of lipid-anchored molecules by high-speed single particle tracking and correlated positions of membrane molecules with the nanoscopic organization of the AIS cytoskeleton.We observe a strong reduction in mobility early in AIS development. Membrane protein motion in the AIS plasma membrane is confined to a repetitive pattern of ~190 nm spaced segments along the AIS axis as early as DIV4 and this pattern alternates with actin rings. Our data provide a new model for the mechanism of the AIS diffusion barrier.


Author(s):  
E.D. Wolf

Most microelectronics devices and circuits operate faster, consume less power, execute more functions and cost less per circuit function when the feature-sizes internal to the devices and circuits are made smaller. This is part of the stimulus for the Very High-Speed Integrated Circuits (VHSIC) program. There is also a need for smaller, more sensitive sensors in a wide range of disciplines that includes electrochemistry, neurophysiology and ultra-high pressure solid state research. There is often fundamental new science (and sometimes new technology) to be revealed (and used) when a basic parameter such as size is extended to new dimensions, as is evident at the two extremes of smallness and largeness, high energy particle physics and cosmology, respectively. However, there is also a very important intermediate domain of size that spans from the diameter of a small cluster of atoms up to near one micrometer which may also have just as profound effects on society as “big” physics.


Author(s):  
N. Yoshimura ◽  
K. Shirota ◽  
T. Etoh

One of the most important requirements for a high-performance EM, especially an analytical EM using a fine beam probe, is to prevent specimen contamination by providing a clean high vacuum in the vicinity of the specimen. However, in almost all commercial EMs, the pressure in the vicinity of the specimen under observation is usually more than ten times higher than the pressure measured at the punping line. The EM column inevitably requires the use of greased Viton O-rings for fine movement, and specimens and films need to be exchanged frequently and several attachments may also be exchanged. For these reasons, a high speed pumping system, as well as a clean vacuum system, is now required. A newly developed electron microscope, the JEM-100CX features clean high vacuum in the vicinity of the specimen, realized by the use of a CASCADE type diffusion pump system which has been essentially improved over its predeces- sorD employed on the JEM-100C.


Author(s):  
William Krakow

In the past few years on-line digital television frame store devices coupled to computers have been employed to attempt to measure the microscope parameters of defocus and astigmatism. The ultimate goal of such tasks is to fully adjust the operating parameters of the microscope and obtain an optimum image for viewing in terms of its information content. The initial approach to this problem, for high resolution TEM imaging, was to obtain the power spectrum from the Fourier transform of an image, find the contrast transfer function oscillation maxima, and subsequently correct the image. This technique requires a fast computer, a direct memory access device and even an array processor to accomplish these tasks on limited size arrays in a few seconds per image. It is not clear that the power spectrum could be used for more than defocus correction since the correction of astigmatism is a formidable problem of pattern recognition.


Author(s):  
C. O. Jung ◽  
S. J. Krause ◽  
S.R. Wilson

Silicon-on-insulator (SOI) structures have excellent potential for future use in radiation hardened and high speed integrated circuits. For device fabrication in SOI material a high quality superficial Si layer above a buried oxide layer is required. Recently, Celler et al. reported that post-implantation annealing of oxygen implanted SOI at very high temperatures would eliminate virtually all defects and precipiates in the superficial Si layer. In this work we are reporting on the effect of three different post implantation annealing cycles on the structure of oxygen implanted SOI samples which were implanted under the same conditions.


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