Structure retention of silica gel-encapsulated bacteriorhodopsin in purple membrane and in lipid nanodiscs

Quantitative structure-retention relationships for a series of 30 mixed ?-diketonato complexes of cobalt(III), chromium(III) and ruthenium(III) were derived by multiple linear regression analyses using molecular descriptors obtained by quantum chemical calculations. The retention parameters were obtained by thin layer chromatography on silica gel using mono and two-component solvent systems. The molecular descriptors included in the multiple linear regression analysis were molecular weight, molecular volume, surface area, hydrophilic-lipophilic balance, percent hydrophilic surface area, dipole moment, polarizability, refractivity, energy of the highest occupied molecular orbital and energy of the lowest unoccupied molecular orbital. High agreement between the experimental and predicted retention parameters was obtained when polarizability and the hydrophilic-lipophilic balance were used as the molecular descriptors. Comparison of the models with those established on polyacrylonitrile showed that the structure of the sorbent is responsible for the chromatographic behaviour of the same compounds. The presented models can be used for the prediction of the retention of new solutes in screening chromatographic systems.

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Photoreceptor disk membrane substructures by means of the complementary freeze-fracture-replica methods

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100081644 ◽

1978 ◽

Vol 36 (2) ◽

pp. 156-157

Author(s):

A. Tonosaki ◽

M. Yamasaki ◽

H. Washioka ◽

J. Mizoguchi

Keyword(s):

Cell Membranes ◽

Freeze Fracture ◽

Purple Membrane ◽

Remarkable Case ◽

Single Face ◽

Disk Membrane ◽

Associated Proteins ◽

Photoreceptor Membranes

A vertebrate disk membrane is composed of 40 % lipids and 60 % proteins. Its fracture faces have been classed into the plasmic (PF) and exoplasmic faces (EF), complementary with each other, like those of most other types of cell membranes. The hypothesis assuming the PF particles as representing membrane-associated proteins has been challenged by serious questions if they in fact emerge from the crystalline formation or decoration effects during freezing and shadowing processes. This problem seems to be yet unanswered, despite the remarkable case of the purple membrane of Halobacterium, partly because most observations have been made on the replicas from a single face of specimen, and partly because, in the case of photoreceptor membranes, the conformation of a rhodopsin and its relatives remains yet uncertain. The former defect seems to be partially fulfilled with complementary replica methods.

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Difference Fourier Analysis of Glucose Embedded and Frozen Hydrated Purple Membrane

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100053164 ◽

1982 ◽

Vol 40 ◽

pp. 74-75

Author(s):

Jules S. Jaffe ◽

Robert M. Glaeser

Keyword(s):

Purple Membrane ◽

Data Set ◽

High Resolution Data ◽

X Ray ◽

X Ray Crystallography ◽

Fourier Techniques ◽

Versus Protein ◽

The Difference ◽

Difference Fourier ◽

Ideal Method

Although difference Fourier techniques are standard in X-ray crystallography it has only been very recently that electron crystallographers have been able to take advantage of this method. We have combined a high resolution data set for frozen glucose embedded Purple Membrane (PM) with a data set collected from PM prepared in the frozen hydrated state in order to visualize any differences in structure due to the different methods of preparation. The increased contrast between protein-ice versus protein-glucose may prove to be an advantage of the frozen hydrated technique for visualizing those parts of bacteriorhodopsin that are embedded in glucose. In addition, surface groups of the protein may be disordered in glucose and ordered in the frozen state. The sensitivity of the difference Fourier technique to small changes in structure provides an ideal method for testing this hypothesis.

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Fresnel interference in point-projection imaging of purple membrane

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100140609 ◽

1995 ◽

Vol 53 ◽

pp. 846-847

Author(s):

X. Zhang ◽

J. Spence ◽

W. Qian ◽

D. Taylor ◽

K. Taylor

Keyword(s):

Detection Efficiency ◽

Mean Free Path ◽

Purple Membrane ◽

Unit Cell Dimension ◽

Experimental Point ◽

Membrane Thickness ◽

Low Energies ◽

Channel Plate ◽

Point Projection ◽

Inelastic Mean Free Path

Experimental point-projection shadow microscope (PPM) images of uncoated, unstained purple membrane (PM, bacteriorhodopsin, a membrane protein from Halobacterium holobium) were obtained recently using 100 volt electrons. The membrane thickness is about 5 nm and the hexagonal unit cell dimension 6 nm. The images show contrast around the edges of small holes, as shown in figure 1. The interior of the film is opaque. Since the inelastic mean free path for 100V electrons in carbon (about 6 Å) is much less than the sample thickness, the question arises that how much, if any, transmission of elastically scattered electrons occurs. A large inelastic contribution is also expected, attenuated by the reduced detection efficiency of the channel plate at low energies. Quantitative experiments using an energy-loss spectrometer are planned. Recently Shedd has shown that at about 100V contrast in PPM images of thin gold films can be explained as Fresnel interference effects between different pinholes in the film, separated by less than the coherence width.

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