Robust High-Yield Methodologies for 2H and 2H/15N/13C Labeling of Proteins for Structural Investigations Using Neutron Scattering and NMR

2015 ◽  
pp. 3-25 ◽  
Author(s):  
Anthony P. Duff ◽  
Karyn L. Wilde ◽  
Agata Rekas ◽  
Vanessa Lake ◽  
Peter J. Holden
Keyword(s):  
Neutron Scattering ◽  
High Yield ◽  
Structural Investigations ◽  
13C Labeling

Structural Investigations of Protein–Lipid Complexes Using Neutron Scattering

2019 ◽  
pp. 201-251 ◽  
Author(s):  
Luke A. Clifton ◽  
Stephen C. L. Hall ◽  
Najet Mahmoudi ◽  
Timothy J. Knowles ◽  
Frank Heinrich ◽  
...  
Keyword(s):  
Neutron Scattering ◽  
Structural Investigations

scholarly journals Small-Angle Neutron Scattering Study of Bicelles and Proteobicelles with Incorporated Mitochondrial Cytochrome c Oxidase

2020 ◽  
Vol 65 (8) ◽  
pp. 662
Author(s):  
K. Siposova ◽  
V. I. Petrenko ◽  
O. I. Ivankov ◽  
L. A. Bulavin ◽  
A. Musatov
Keyword(s):  
Cytochrome C ◽  
Neutron Scattering ◽  
Cytochrome C Oxidase ◽  
Small Angle ◽  
Small Angle Neutron Scattering ◽  
Membrane System ◽  
Aggregation State ◽  
Structural Investigations ◽  
Neutron Scattering Study ◽  
Average Size

The structural investigations of a model membrane system, bicelles, and the aggregation state of isolated and purified bovine heart cytochrome c oxidase (CcO) in bicelles have been performed using small-angle neutron scattering (SANS), SANS contrast variation, and complemented by various biophysical and biochemical techniques. The average size of bicelles prepared from long-chain 1,2-dimyristoyl-sn-glycero-3-phosphocholine and short-chain 1,2-dihexanoyl-sn-glycero-3-phosphocholine was found to be about 22 nm with a thickness of about 4 nm. Enzyme in bicelles was remained active and structurally unaltered. The estimated volume of protein in bicelles of 240 nm3 corresponded well to the monomeric form of CcO. The ab initio modeling supports the experimental data and suggests that CcO in bicelles is a homogeneous monomeric complex incorporated into bicelles.

Structural investigations of La0.8Sr0.2CrO3 by X-ray and neutron scattering

2009 ◽  
Vol 475 (1-2) ◽  
pp. 614-618 ◽  
Author(s):  
A.K. Patra ◽  
Sathi Nair ◽  
P.U. Sastry ◽  
A.K. Tyagi
Keyword(s):  
Neutron Scattering ◽  
Structural Investigations ◽  
X Ray

Observation of biological macromolecules using various electron microscopic methods

1978 ◽  
Vol 36 (2) ◽  
pp. 170-171
Author(s):  
Mitsuo Ohtsuki ◽  
Michael Sogard
Keyword(s):  
Electron Microscope ◽  
Phase Contrast ◽  
Image Interpretation ◽  
Density Lipoprotein ◽  
Biological Macromolecules ◽  
Contrast Effects ◽  
Biological Structure ◽  
Electron Microscopic ◽  
Structural Investigations ◽  
Staining Techniques

Structural investigations of biological macromolecules commonly employ CTEM with negative staining techniques. Difficulties in valid image interpretation arise, however, due to problems such as variability in thickness and degree of penetration of the staining agent, noise from the supporting film, and artifacts from defocus phase contrast effects. In order to determine the effects of these variables on biological structure, as seen by the electron microscope, negative stained macromolecules of high density lipoprotein-3 (HDL3) from human serum were analyzed with both CTEM and STEM, and results were then compared with CTEM micrographs of freeze-etched HDL3. In addition, we altered the structure of this molecule by digesting away its phospholipid component with phospholipase A2 and look for consistent changes in structure.

Quality films from carbon fiber evaporation for thorough coverage of TEM grids

1988 ◽  
Vol 46 ◽  
pp. 418-419
Author(s):  
N. Tempel ◽  
M. C. Ledbetter
Keyword(s):  
Electron Microscopy ◽  
High Resolution Electron Microscopy ◽  
Carbon Films ◽  
Low Noise ◽  
Vacuum Evaporation ◽  
High Yield ◽  
Good Adhesion ◽  
Noise Structure ◽  
Resolution Electron ◽  
Carbon Foils

Carbon films have been a support of choice for high resolution electron microscopy since the introduction of vacuum evaporation of carbon. The desirable qualities of carbon films and methods of producing them has been extensively reviewed. It is difficult to get a high yield of grids by many of these methods, especially if virtually all of the windows must be covered with a tightly bonded, quality film of predictable thickness. We report here a method for producing carbon foils designed to maximize these attributes: 1) coverage of virtually all grid windows, 2) freedom from holes, wrinkles or folds, 3) good adhesion between film and grid, 4) uniformity of film and low noise structure, 5) predictability of film thickness, and 6) reproducibility.Our method utilizes vacuum evaporation of carbon from a fiber onto celloidin film and grid bars, adhesion of the film complex to the grid by carbon-carbon contact, and removal of the celloidin by acetone dissolution. Materials must be of high purity, and cleanliness must be rigorously maintained.

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