Ultracentrifuge Studies with Rayleigh Interference Optics.I. General Application

Richards, E. G.; Schachman, H. K.

doi:10.1021/j150580a008

Cited by 314 publications

(168 citation statements)

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“…The equilibrium speed for al was 6,995 rpm; that for Bs2 was 8,225 rpm. Rayleigh patterns were measured in a Nikon comparator and molecular weights calculated as described by Richards and Schachman (14). The partial specific volume was taken to be 0.705 (15).…”

Section: Methodsmentioning

confidence: 99%

A Biochemical Study of Human Skin Collagen and the Relation between Intra- and Intermolecular Cross-Linking *

Börnstein¹,

Piez²

1964

J. Clin. Invest.

151

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Section: Methodsmentioning

confidence: 99%

A Biochemical Study of Human Skin Collagen and the Relation between Intra- and Intermolecular Cross-Linking *

Börnstein¹,

Piez²

1964

J. Clin. Invest.

151

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“…Any refractive index difference, Δn, between the two solutions contributes to the optical path length so that: ΔY = Δnl/λ, where l is the optical path length and λ is the wavelength of the light source (Richards and Schachman, 1959;Yphantis, 1964). For a non-dialyzable solution component, the refractive index difference is proportional to the refractive index increment: Δn = c(dn/dc) and the extinction coefficient e is replaced by (6) For proteins, dn/dc is relatively independent of composition with an average value of 0.186 ml/g (Huglin, 1972).…”

Section: B Interferencementioning

confidence: 99%

Analytical Ultracentrifugation: Sedimentation Velocity and Sedimentation Equilibrium

Cole

Lary

Moody

et al. 2008

Methods in Cell Biology

368

274

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Analytical ultracentrifugation (AUC) is a versatile and powerful method for the quantitative analysis of macromolecules in solution. AUC has broad applications for the study of biomacromolecules in a wide range of solvents and over a wide range of solute concentrations. Three optical systems are available for the analytical ultracentrifuge (absorbance, interference and fluorescence) that permit precise and selective observation of sedimentation in real time. In particular, the fluorescence system provides a new way to extend the scope of AUC to probe the behavior of biological molecules in complex mixtures and at high solute concentrations. In sedimentation velocity, the movement of solutes in high centrifugal fields is interpreted using hydrodynamic theory to define the size, shape and interactions of macromolecules. Sedimentation equilibrium is a thermodynamic method where equilibrium concentration gradients at lower centrifugal fields are analyzed to define molecule mass, assembly stoichiometry, association constants and solution nonideality. Using specialized sample cells and modern analysis software, researchers can use sedimentation velocity to determine the homogeneity of a sample and define whether it undergoes concentration-dependent association reactions. Subsequently, more thorough model-dependent analysis of velocity and equilibrium experiments can provide a detailed picture of the nature of the species present in solution and their interactions.

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“…Absolute fringe numbers were determined by the white-light fringe method [19]. Measurements were made with a toolmakers micro-comparator, aligned using the air fringes, and were taken at intervals of 0.02 cm radially.…”

Section: Molecular Weight Determinationsmentioning

confidence: 99%

Isolation and Characterisation of Glycoproteins from Sputum

Roberts

1974

European Journal of Biochemistry

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Sputum has been separated into a sol and a gel phase by ultracentrifugation and the components of these phases examined. A glycoprotein of high molecular weight as well as serum albumin, immunoglobulin A, sll acid glycoprotein and lactoferrin were detected in the sol phase by gel filtration and immunodiffusion studies. A glycoprotein of similar chemical composition was isolated from the gel phase by solubilisation in 6 M urea/O. 1 M NaCl followed by fractional precipitation with ethanol.The carbohydrate content of this glycoprotein, termed the bronchial glycoprotein, varied from 58 (w/w) to 69% (w/w) with sputum from different patients and was composed of fucose, galactose, galactosamine, glucosamine and N-acetylneuraminic acid. Serine, threonine and proline constituted 44% (w/w) of the amino acid residues of the bronchial glycoprotein and an alkaline borohydride cleavage study indicated that 0-glycosidic linkages exist between N-acetylgalactosamine and the hydroxyamino acids in the peptide core. Fractionation of the bronchial glycoprotein on DEAESephadex A-25 yielded fractions enriched in sulphate and sialic acid but separate fuco, sialo and sulpho glycoproteins were not obtained. Similarly fractionation of the oligosaccharides released by the action of alkaline borohydride yielded a complex mixture of oligosaccharides but sulphated oligosaccharides free of sialic acid were not obtained. Separation of the oligosaccharides into two fractions of different sizes showed that sulphate content was greatest in the larger oligosaccharides whereas sialic acid content was highest in the smaller oligosaccharides. A methylation study of the bronchial glycoprotein, neuraminidase-digested bronchial glycoprotein and mild acid-hydrolysed bronchial glycoprotein revealed that the N-acetylneuraminic acid was linked to C-3 of penultimate galactose units. Fucose occurs in three different linkages, approximately 50 % is linked to C-2 of penultimate galactose units, approximately 25 % is linked to C-4 of glucosamine residues already substituted at

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Ultracentrifuge Studies with Rayleigh Interference Optics.I. General Application

Cited by 314 publications

References 1 publication

A Biochemical Study of Human Skin Collagen and the Relation between Intra- and Intermolecular Cross-Linking *

A Biochemical Study of Human Skin Collagen and the Relation between Intra- and Intermolecular Cross-Linking *

Analytical Ultracentrifugation: Sedimentation Velocity and Sedimentation Equilibrium

Isolation and Characterisation of Glycoproteins from Sputum

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