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Biomedical subjects

O H Griffith

Publications and source records attributed to O H Griffith.

At least 109 records · Page 6Linked to original sources
PubMed1975

Bilayer structure in phospholipid-cytochrome c model membranes.

Lipid protein interactions in biological membranes differ markedly depending on whether the protein is intrinsic or extrinsic. These interactions are studied using lipid spin labels diffused into model systems consisting of phospholipid bilayers and a specific protein. Recently, an intrinsic protein complex, cytochrome oxidase, was examined and the data suggest there is a boundary layer of immobilized lipid between the hydrophobic protein surfaces and adjacent fluid bilayer regions. In the present study, a typical extrinsic protein, cytochrome c, was complexed with a cardiolipin/lecithin (1:4 by weight) mixture. The phospholipids in the presence and absence of cytochrome c exhibit typical bilayer behavior as jedged by four spin-labeling criteria: fluidity gradient, spectral anisotropy of oriented bilayers, response to hydration and the polarity profile. Any effects of cytochrome c on the ESR spectra of lipid spin labels are small, in contrast to the effects of intrinsic proteins. These data are consistent with electrostatic binding of cytochrome c to the charged groups of the phospholipids, and indicate that the presence of extrinsic proteins will not interfere with measurements of boundary lipid in intact biological membranes.

Animals↗
PubMed1974

Photoelectron quantum yields of the amino acids.

The photoelectron quantum yields of 21 common amino acids and 15 polyamino acids were measured in the 180-240 nm wavelength region. On the average, the quantum yields of these two groups exhibit quite similar wavelength dependence. For lambda > 220 nm all amino acid and polyamino acid quantum yields are </=10(-7) electrons/(incident) photon. The mean yields increase to about 5 x 10(-7) electrons/photon at 200 nm and 5 x 10(-6) electrons/photon at 180 nm. L-tryptophan, L-tyrosine, and poly-L-tryptophan exhibit above average yields between 180 and 200 nm. Comparison with the dye phthalocyanine indicates that the quantum yield of the dye is two orders of magnitude greater than that of the amino acids from 200 to 240 nm, suggesting the feasibility of photoelectron labeling studies of biological surfaces.

Amino Acids↗
PubMed1974

Photoelectron quantum yields of hemin, hemoglobin, and apohemoglobin. Possible applications to photoelectron microscopy of heme proteins in biological membranes.

Hemoglobin is examined as a model system for intrinsic photoelectron labeling experiments. The absolute photoelectron quantum yields of hemin, hemoglobin, and apohemoglobin thin films were measured in the 180-230 nm wavelength region. Hemin exhibits a quantum yield of approximately 6 x 10(-4) electrons per incident photon at 180 nm, 9 x 10(-5) electrons per incident photon at 210 nm, and 2 x 10(-6) electrons per incident photon at 230 nm. At all wavelengths the hemin curve lies approximately a factor of 20 above that of hemoglobin and two orders of magnitude above that of apohemoglobin. High image contrast is observed between hemin and apohemoglobin in low magnification photoelectron micrographs, suggesting the feasibility of intrinsic labeling studies involving heme proteins. The quantum yield of hemoglobin is discussed in terms of linear contributions from heme groups and protein weighted by their relative surface areas. The fractional surface areas based on the known structure of hemoglobin are consistent with values derived from the quantum yields of hemin and apohemoglobin.

Animals↗
PubMed1974

Lipid binding to the amphipathic membrane protein cytochrome b5.

The lipid binding properties of the membrane protein cytochrome b(5) (detergent-extracted from calf liver microsomal preparations) were characterized by studying the interaction of spin-labeled lipids (5-, 12-, and 16-doxylstearic acid and 5- and 16-doxylphosphatidyl-choline, where doxyl refers to the nitroxide moiety) with cytochrome b(5), using electron spin resonance spectroscopy. The intact cytochrome b(5) molecule immobilizes all of the lipid spin labels, while the segment of cytochrome b(5) released by trypsin does not affect lipid mobility. The immobilization of lipid spin labels on the hydrophobic surface of intact cytochrome b(5) is not appreciably altered by associating the protein with liposomes. Differences in polarity of the lipid binding sites between cytochrome b(5) and phospholipid vesicles were also observed. The lipid binding sites on cytochrome b(5) are hydrophobic by conventional criteria, but are more polar than the interior of fluid phospholipid bilayers.

Animals↗
PubMed1973

Contrast in the photoelectric effect of organic and biochemical surfaces. A first step towards selective labeling in photoelectron microscopy.

The photoelectric effect can provide the physical basis for a new method of mapping organic and biological surfaces. The technique, photoelectron microscopy, is similar to fluorescence microscopy using incident ultraviolet light except that photoejected electrons form the image of the specimen surface. In this work the minimum wavelengths of incident light required to produce an image were determined for the molecules 3,6-bis(dimethylamino)acridine (acridine orange) (I), benzo[a]pyrene (II), N,N,N',N'-tetraphenylbenzidine (III), and copper phthalocyanine (IV). The photoelectron image thresholds for these compounds are 220 (I), 215 (II), 220 (III), and 240 nm (IV), all +/-5 nm. Contrast of I-IV with respect to typical protein, lipid, nucleic acid, and polysaccharide surfaces was examined over the wavelength range 240-180 nm. The low magnification micrographs exhibited bright areas corresponding to I-IV but dark regions for the biochemical surfaces. The high contrast suggests the feasibility of performing extrinsic photoelectron microscopy experiments through selective labeling of sites on biological surfaces.

Acridines↗
PubMed1973

Evidence for boundary lipid in membranes.

Cytochrome oxidase (EC 1.9.3.1) isolated from beef-heart mitochondria with an appropriate phospholipid content forms vesicular structures. Lipid-protein interactions in this model membrane system were studied with the lipid spin label, 16-doxylstearic acid. As the phospholipid/protein ratio is varied, two spectral components are observed. At low phospholipid/protein ratios (</=0.19 mg of phospholipid per mg of protein) the lipid spin label is highly immobilized. At higher phospholipid content an additional component characteristic of fluid lipid bilayers is evident. By summation of digitalized spectra and subsequent integration it was shown that all composite spectra could be approximated by assuming only two components are present, and that the amount of phospholipid bound to the protein is independent of the extent of the fluid bilayer region. The experimentally determined amount of phospholipid for maximum occupancy of protein-bound sites is about 0.2 mg of phospholipid per 1.0 mg of protein. Calculations show that this ratio is consistent with a single layer of phospholipid surrounding the protein complex. The data are interpreted as evidence for a boundary of immobilized lipid between the hydrophobic protein and adjacent fluid bilayer regions in this membrane model system.

Animals↗
PubMed1972

Photoelectron microscopy: a new approach to mapping organic and biological surfaces.

A general method of imaging organic and biological surfaces based on the photoelectric effect is reported. For the experiments, a photoelectron emission microscope was constructed. It is an ultrahigh vacuum instrument using electrostatic electron lenses, microchannel plate image intensifier, cold stage, hydrogen excitation source, and magnesium fluoride optics. The organic surfaces examined were grid patterns of acridine orange, fluorescein, and benzo(a)pyrene on a Butvar surface. A biological sample, sectioned rat epididymis, was also imaged by the new photoelectron microscope. Good contrast was obtained in these initial low magnification experiments. These data demonstrate the feasibility of mapping biological surfaces according to differences in ionization potentials of exposed molecules. A number of technical difficulties, such as the intensity of the excitation source, must be solved before high resolution experiments are practical. However, it is probable that this approach can be useful, even at low magnifications, in determination of the properties of organic and biological surfaces.

Acridines↗