Biomolecular analysis of elastic fibre molecules.
Elastic fibres are macromolecular extracellular matrix assemblies that endow dynamic connective tissues such as arteries, lungs and skin with the property of elastic recoil. Here, we describe how we have purified elastic fibre molecules and then analysed them using a range of biochemical and biomolecular approaches. Such approaches have provided powerful insights into the complex hierarchical processes of extracellular matrix assembly. We outline molecular interaction and kinetics assays using Biacore, biophysical approaches such as multi-angle laser light scattering and analytical ultracentrifugation which provide information on molecular and macromolecular shape and mass in solution, the visualisation of molecules and assemblies using microscopy approaches such as atomic force microscopy and environmental scanning electron microscopy, and compositional analysis of macromolecular complexes using mass spectrometry. Data from these in vitro analytical approaches can be combined to develop powerful new models of elastic fibre assembly.
Cain
Stuart A
SA
Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Michael Smith Building, Oxford Road, Manchester, UK.
Raynal
Bertrand
B
Hodson
Nigel
N
Shuttleworth
Adrian
A
Kielty
Cay M
CM
D000818:Animals; D004547:Elastic Tissue / Q000737:chemistry*; D000071837:Fibrillins; D006801:Humans; D008027:Light; D008840:Microfilament Proteins / Q000302:isolation & purification*; D018625:Microscopy, Atomic Force; D019906:Nuclear Magnetic Resonance, Biomolecular; D012542:Scattering, Radiation; D013499:Surface Properties; D014461:Ultracentrifugation
D000071837:Fibrillins; D008840:Microfilament Proteins