Origin of the Surprising Mechanical Stability of Kinesin’s Neck Coiled Coil

2021 ◽  
Vol 17 (2) ◽  
pp. 1017-1029
Author(s):  
Shu-Xia Liu ◽  
Gang Lü ◽  
Hui Zhang ◽  
Yi-Zhao Geng ◽  
Qing Ji
Keyword(s):  
Mechanical Stability ◽  
Coiled Coil

scholarly journals Structural Determinants of Coiled Coil Mechanics

2019 ◽  
Author(s):  
Patricia Lopez-Garcia ◽  
Melis Goktas ◽  
Ana E. Bergues-Pupo ◽  
Beate Koksch ◽  
Daniel Varon Silva ◽  
...  
Keyword(s):  
Transition State ◽  
Single Molecule ◽  
Mechanical Stability ◽  
Coiled Coil ◽  
Building Blocks ◽  
Shear Loading ◽  
Hydrophobic Core ◽  
Structural Determinants ◽  
Helix Propensity ◽  
Core Packing

The natural abundance of coiled coil (CC) motifs in the cytoskeleton and the extracellular matrix suggests that CCs play a crucial role in the bidirectional mechanobiochemical signaling between cells and the matrix. Their functional importance and structural simplicity has allowed the development of numerous applications, such as protein-origami structures, drug delivery systems and biomaterials. With the goal of establishing CCs as nanomechanical building blocks, we investigated the importance of helix propensity and hydrophobic core packing on the mechanical stability of 4-heptad CC heterodimers. Using single-molecule force spectroscopy, we show that both parameters determine the force-induced dissociation in shear loading geometry; however, with different effects on the energy landscape. Decreasing the helix propensity lowers the transition barrier height, leading to a concomitant decrease in the distance to the transition state. In contrast, a less tightly packed hydrophobic core increases the distance to the transition state. We propose that this sequence-structure-mechanics relationship is evolutionarily optimized in natural CCs and can be used for tuning their mechanical properties in applications.

scholarly journals Single molecule mechanics of the kinesin neck

2009 ◽  
Vol 106 (17) ◽  
pp. 6992-6997 ◽  
Author(s):  
Thomas Bornschlögl ◽  
Günther Woehlke ◽  
Matthias Rief
Keyword(s):  
Single Molecule ◽  
Leucine Zipper ◽  
Structural Integrity ◽  
Mechanical Stability ◽  
Molecular Motor ◽  
Coiled Coil ◽  
Force Microscopy ◽  
Atomic Force ◽  
Kinetics Of ◽  
High Mechanical Stability

Structural integrity as well as mechanical stability of the parts of a molecular motor are crucial for its function. In this study, we used high-resolution force spectroscopy by atomic force microscopy to investigate the force-dependent opening kinetics of the neck coiled coil of Kinesin-1 from Drosophila melanogaster. We find that even though the overall thermodynamic stability of the neck is low, the average opening force of the coiled coil is >11 pN when stretched with pulling velocities >150 nm/s. These high unzipping forces ensure structural integrity during motor motion. The high mechanical stability is achieved through a very narrow N-terminal unfolding barrier if compared with a conventional leucine zipper. The experimentally mapped mechanical unzipping profile allows direct assignment of distinct mechanical stabilities to the different coiled-coil subunits. The coiled-coil sequence seems to be tuned in an optimal way to ensure both mechanical stability as well as motor regulation through charged residues.

scholarly journals Tuning coiled coil stability with histidine-metal coordination

Nanoscale ◽  
2018 ◽  
Vol 10 (48) ◽  
pp. 22725-22729 ◽  
Author(s):  
Isabell Tunn ◽  
Alberto S. de Léon ◽  
Kerstin G. Blank ◽  
Matthew J. Harrington
Keyword(s):  
Mechanical Stability ◽  
Coiled Coil ◽  
Coiled Coils ◽  
Metal Coordination

Reinforcing coiled coils with histidine-metal coordination reversibly increases their thermodynamic and mechanical stability with implications for biomimetic hydrogel design.

An improved design for an Scanning Tunnelling Microscope (STM) for use in a Transmission Electron Microscope (TEM)

1991 ◽  
Vol 49 ◽  
pp. 384-385
Author(s):  
W.K. Lo ◽  
J.C.H. Spence
Keyword(s):  
Electron Microscope ◽  
Transmission Electron Microscope ◽  
Mechanical Stability ◽  
Scanning Tunnelling Microscope ◽  
Reflection Mode ◽  
Specimen Holder ◽  
Transmission Electron ◽  
Scanning Tunnelling ◽  
Improved Design

An improved design for a combination Scanning Tunnelling Microscope/TEM specimen holder is presented. It is based on earlier versions which have been used to test the usefulness of such a device. As with the earlier versions, this holder is meant to replace the standard double-tilt specimen holder of an unmodified Philips 400T TEM. It allows the sample to be imaged simultaneously by both the STM and the TEM when the TEM is operated in the reflection mode (see figure 1).The resolution of a STM is determined by its tip radii as well as its stability. This places strict limitations on the mechanical stability of the tip with respect to the sample. In this STM the piezoelectric tube scanner is rigidly mounted inside the endcap of the STM holder. The tip coarse approach to the sample (z-direction) is provided by an Inchworm which is located outside the TEM vacuum.

Experiments with a scanning tunneling microscope (STM) mounted in a scanning electron microscope (SEM)

1986 ◽  
Vol 44 ◽  
pp. 636-639
Author(s):  
Oliver C. Wells ◽  
Mark E. Welland
Keyword(s):  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
Mechanical Stability ◽  
Tunneling Current ◽  
Feedback System ◽  
Scanning Tunneling ◽  
Surface Profiling ◽  
Close Proximity ◽  
Metal Tip ◽  
Scanning Electron

Scanning tunneling microscopes (STM) exist in two versions. In both of these, a pointed metal tip is scanned in close proximity to the specimen surface by means of three piezos. The distance of the tip from the sample is controlled by a feedback system to give a constant tunneling current between the tip and the sample. In the low-end STM, the system has a mechanical stability and a noise level to give a vertical resolution of between 0.1 nm and 1.0 nm. The atomic resolution STM can show individual atoms on the surface of the specimen.A low-end STM has been put into the specimen chamber of a scanning electron microscope (SEM). The first objective was to investigate technological problems such as surface profiling. The second objective was for exploratory studies. This second objective has already been achieved by showing that the STM can be used to study trapping sites in SiO2.

Electron microscopy of keratinocytes cultured on a collagen substrate used as an allograft for the treatment of chronic wounds

1992 ◽  
Vol 50 (2) ◽  
pp. 1104-1105
Author(s):  
Debby A. Jennings ◽  
Michael J. Morykwas ◽  
Louis C. Argenta
Keyword(s):  
Electron Microscopy ◽  
Cell Suspension ◽  
Single Cell ◽  
Chronic Wounds ◽  
Mechanical Stability ◽  
Uv Light ◽  
Type I ◽  
Single Cell Suspension ◽  
Collagen Substrate ◽  
Dermal Collagen

Grafts of cultured allogenic or autogenic keratlnocytes have proven to be an effective treatment of chronic wounds and burns. This study utilized a collagen substrate for keratinocyte and fibroblast attachment. The substrate provided mechanical stability and augmented graft manipulation onto the wound bed. Graft integrity was confirmed by light and transmission electron microscopy.Bovine Type I dermal collagen sheets (100 μm thick) were crosslinked with 254 nm UV light (13.5 Joules/cm2) to improve mechanical properties and reduce degradation. A single cell suspension of third passage neonatal foreskin fibroblasts were plated onto the collagen. Five days later, a single cell suspension of first passage neonatal foreskin keratinocytes were plated on the opposite side of the collagen. The grafts were cultured for one month.The grafts were fixed in phosphate buffered 4% formaldehyde/1% glutaraldehyde for 24 hours. Graft pieces were then washed in 0.13 M phosphate buffer, post-fixed in 1% osmium tetroxide, dehydrated, and embedded in Polybed 812.

P20 phosphor on polyimide as a large area high-resolution transmission screen for slow scan CCD systems

1995 ◽  
Vol 53 ◽  
pp. 20-21
Author(s):  
C. C. Ahn ◽  
S. Karnes ◽  
M. Lvovsky ◽  
C. M. Garland ◽  
H. A. Atwater ◽  
...  
Keyword(s):  
Mechanical Stability ◽  
High Energy ◽  
Practical Implementation ◽  
Line Pair ◽  
Modulation Transfer ◽  
Large Area ◽  
Ccd Imaging ◽  
Transmission Electron ◽  
The One ◽  
Beam Broadening

The bane of CCD imaging systems for transmission electron microscopy at intermediate and high voltages has been their relatively poor modulation transfer function (MTF), or line pair resolution. The problem originates primarily with the phosphor screen. On the one hand, screens should be thick so that as many incident electrons as possible are converted to photons, yielding a high detective quantum efficiency(DQE). The MTF diminishes as a function of scintillator thickness however, and to some extent as a function of fluorescence within the scintillator substrates. Fan has noted that the use of a thin layer of phosphor beneath a self supporting 2μ, thick Al substrate might provide the most appropriate compromise for high DQE and MTF in transmission electron microcscopes which operate at higher voltages. Monte Carlo simulations of high energy electron trajectories reveal that only little beam broadening occurs within this thickness of Al film. Consequently, the MTF is limited predominantly by broadening within the thin phosphor underlayer. There are difficulties however, in the practical implementation of this design, associated mostly with the mechanical stability of the Al support film.

A conduction-cooled stage for high-resolution Cryo-SEM

1992 ◽  
Vol 50 (2) ◽  
pp. 1282-1283
Author(s):  
John G. Sheehan
Keyword(s):  
High Resolution ◽  
Liquid Nitrogen ◽  
Mechanical Stability ◽  
Cooling System ◽  
Cold Trap ◽  
Nitrogen Gas ◽  
Particulate Suspensions ◽  
Advantages And Disadvantages ◽  
Convection Cooling ◽  
Styrene Butadiene

The goal is to examine with high resolution cryo-SEM aqueous particulate suspensions used in coatings for printable paper. A metal-coating chamber for cryo-preparation of such suspensions was described previously. Here, a new conduction-cooling system for the stage and cold-trap in an SEM specimen chamber is described. Its advantages and disadvantages are compared to a convection-cooling system made by Hexland (model CT1000A) and its mechanical stability is demonstrated by examining a sample of styrene-butadiene latex.In recent high resolution cryo-SEM, some stages are cooled by conduction, others by convection. In the latter, heat is convected from the specimen stage by cold nitrogen gas from a liquid-nitrogen cooled evaporative heat exchanger. The advantage is the fast cooling: the Hexland CT1000A cools the stage from ambient temperature to 88 K in about 20 min. However it consumes huge amounts of liquid-nitrogen and nitrogen gas: about 1 ℓ/h of liquid-nitrogen and 400 gm/h of nitrogen gas. Its liquid-nitrogen vessel must be re-filled at least every 40 min.

TMCC3 localizes at the three-way junctions for the proper tubular network of the endoplasmic reticulum

2019 ◽  
Vol 476 (21) ◽  
pp. 3241-3260
Author(s):  
Sindhu Wisesa ◽  
Yasunori Yamamoto ◽  
Toshiaki Sakisaka
Keyword(s):  
Endoplasmic Reticulum ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Mammalian Cells ◽  
Coiled Coil ◽  
Transmembrane Domains ◽  
Domain Family ◽  
Coiled Coil Domain ◽  
U2os Cells ◽  
Er Membrane

The tubular network of the endoplasmic reticulum (ER) is formed by connecting ER tubules through three-way junctions. Two classes of the conserved ER membrane proteins, atlastins and lunapark, have been shown to reside at the three-way junctions so far and be involved in the generation and stabilization of the three-way junctions. In this study, we report TMCC3 (transmembrane and coiled-coil domain family 3), a member of the TEX28 family, as another ER membrane protein that resides at the three-way junctions in mammalian cells. When the TEX28 family members were transfected into U2OS cells, TMCC3 specifically localized at the three-way junctions in the peripheral ER. TMCC3 bound to atlastins through the C-terminal transmembrane domains. A TMCC3 mutant lacking the N-terminal coiled-coil domain abolished localization to the three-way junctions, suggesting that TMCC3 localized independently of binding to atlastins. TMCC3 knockdown caused a decrease in the number of three-way junctions and expansion of ER sheets, leading to a reduction of the tubular ER network in U2OS cells. The TMCC3 knockdown phenotype was partially rescued by the overexpression of atlastin-2, suggesting that TMCC3 knockdown would decrease the activity of atlastins. These results indicate that TMCC3 localizes at the three-way junctions for the proper tubular ER network.

SiO2 Entrapment of Animal Cells for Hybrid Bioartificial Organs

2000 ◽  
Vol 628 ◽  
Author(s):  
Giovanni Carturan ◽  
Renzo Dal Monte ◽  
Maurizio Muraca
Keyword(s):  
Gas Phase ◽  
Mechanical Stability ◽  
Porous Silica ◽  
Collagen Matrix ◽  
Homogeneous Layer ◽  
Animal Cells ◽  
Bioartificial Organs ◽  
Liver And Pancreas ◽  
Encapsulation Method ◽  
Metabolic Activities

ABSTRACTSi-alkoxides in gas phase are reactive towards the surface of animal cells, depositing a homogeneous layer of porous silica. This encapsulation method preserves cell viability and does not alter the hindrance of the biological load.In the prospective use for the design of a hybrid bioartificial liver, hepatocytes in a collagen matrix can be entrapped by the siliceous deposit which provides definite mechanical stability to the collagen matrix and molecular cutoff vs. high molecular weight proteins, including immunoglobulins. The functionality of the encapsulated cell load is maintained for the expressions of typical liver and pancreas metabolic activities.

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