[10] Isolation and characterization of insoluble and soluble elastins

Soskel, Norman T.; Wolt, Terril B.; Sandberg, Lawrence B.

doi:10.1016/0076-6879(87)44179-7

Cited by 28 publications

(25 citation statements)

References 32 publications

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“…Elastin was isolated by four standard methods after dissecting the tissue free from adhering fats: (i) The Lansing procedure (heating at 958C in 0.1M NaOH for 45 min), 30 (ii) cyanogen bromide extraction (shaking for 24 hr at room temperature in CNBr at a concentration of 50 mg/ml in 70% formic acid under nitrogen), 31 and (iii) collagenase digestion, 31 with or without DTE to preserve microfibrillar components. 32 aelastin was prepared by hydrolysis of aortic elastin involving repeated boiling and centrifuging in 0.25M oxalic acid.…”

Section: Elastin Preparationmentioning

confidence: 99%

The molecular structure and physical properties of elastin fibers as revealed by Raman microspectroscopy

2008

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Raman microspectroscopy has been used to investigate the structure of alpha-elastin and fibrous elastin from ligament and aorta, and to explore changes associated with mechanical strain and temperature. Although no vibrational modes associated with cross-linking of the fibers could be identified, the secondary structure of dehydrated fibrous elastin was significantly different from alpha-elastin. The former differed from previous experimental measurements, but was close to the theoretical predictions with 36% beta-structures, 46% unordered, and 18% alpha-helix. Alpha-elastin contained 29% beta-structures, 53% unordered, and 18% alpha-helix. In nuchal fibers the amide I mode was polarized, consistent with the peptide bond. Strains of up to 60% in ligament fiber bundles resulted in no significant shifts in peak position or in secondary structure. Polarization measurements revealed that the peptide bonds and several side chains re-orientated closer to the fiber axis. Heating nuchal fibers to 60 degrees C to increase the energetic component of the elasticity was associated with a 30% increase in the proportion of beta-structures in the amide I band, a 50% increase in the amide III band, and a 50% reduction in the signal from bound water.

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Section: Elastin Preparationmentioning

confidence: 99%

The molecular structure and physical properties of elastin fibers as revealed by Raman microspectroscopy

2008

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“…The incident (k i ) light from the laser passes through a lens, is refracted at the air -sample interface (k and equal to p (c 11 /r), thus providing the value of c 11 . When 33 /r), hence the second elastic coefficient can be determined.…”

Section: Appendix Amentioning

confidence: 99%

“…Elastin was prepared by alkali extraction of bovine nuchal ligament using the Lansing procedure [33]. The ligament consists of roughly parallel arrays of fibres approximately 5 mm in diameter (see below) [34].…”

Section: Fibre and Tissue Samplesmentioning

confidence: 99%

Biomechanics of fibrous proteins of the extracellular matrix studied by Brillouin scattering

Palombo

Winlove

Edginton

et al. 2014

J. R. Soc. Interface.

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Brillouin light scattering (BLS) spectroscopy is a technique that is able to detect thermally excited phonons within a material. The speed of propagation of these phonons can be determined from the magnitude of the Brillouin frequency shift between incident and scattered light, thereby providing a measure of the mechanical properties of the material in the gigahertz range. The mechanical properties of the extracellular matrices of biological tissues and their constituent biopolymers are important for normal tissue function and disturbances in these properties are widely implicated in disease. BLS offers the prospect of measuring mechanical properties on a microscopic scale in living tissues, thereby providing insights into structure-function relationships under normal and pathological conditions. In this study, we investigated BLS in collagen and elastin-the fibrous proteins of the extracellular matrix (ECM). Measurements were made on type I collagen in rat tail tendon, type II collagen in articular cartilage and nuchal ligament elastin. The dependence of the BLS spectrum on fibre orientation was investigated in a backscattering geometry using a reflective substrate. Two peaks, a bulk mode arising from phonon propagation along a quasi-radial direction to the fibre axis and a mode parallel to the surface, depending on sample orientation relative to the fibre axis, could be distinguished. The latter peak was fitted to a model of wave propagation through a hexagonally symmetric elastic solid, and the five components of the elasticity tensor were combined to give axial and transverse Young's, shear and bulk moduli of the fibres. These were 10.2, 8.3, 3.2 and 10.9 GPa, and 6.1, 5.3, 1.9 and 8 GPa for dehydrated type I collagen and elastin, respectively. The former values are close to those previously reported. A microfocused BLS approach was also applied providing selection of single fibres. The moduli of collagen and elastin are much higher than those measured at lower frequency using macroscopic strains, and the difference between them is much less. We therefore believe, like previous investigators, that molecular-scale viscoelastic effects are responsible for the frequency dependence of the fibre biomechanics. Combining BLS with larger-scale mechanical testing methods therefore should, in the future, provide a means of following the evolution of mechanical properties in the formation of the complex structures found in the ECM.

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“…The procedure followed in this study for the isolation of insoluble elastin yields as pure a product as possible 11 by avoiding extremes of temperature and pH adopted in all the other principal methods in current usage for purification of insoluble elastin. Hot alkali treatment, 10 which was once considered the best quality control available in the purification of insoluble elastin, gives rise to protein degradation, 7,12 which compromises studies where structural alterations should be avoided.…”

Section: Discussionmentioning

confidence: 99%

“…The amount of noncollagen, nonelastin (nCnE) material in the vessel wall was then estimated by difference. Estimation of the amount of nCnE-protein still present in the final collagenase-treated elastin preparation not treated with DTE (GC) was made by a procedure previously proposed for calculation of the degree of contamination of elastin by nCnE-proteins 11 as well as by the difference in amino acid composition. The determination was made on specimens from two rings of GC-elastin and two of GC/ DTE-elastin.…”

Section: Biochemical Analysismentioning

confidence: 99%