Isolation and ultrastructural analysis of microfibrillar structures from foetal bovine elastic tissues. Relative abundance and supramolecular architecture of type VI collagen assemblies and fibrillin.
Extensive intact assemblies of matrix macromolecules have been solubilized from foetal calf skin, nuchal ligament and aorta by a new procedure that includes bacterial collagenase digestion under non-reducing, non-denaturing conditions and gel filtration chromatography. Type VI collagen was identified as the major microfibrillar element of these tissues by SDS-PAGE analysis and Western blotting. Rotary shadowing electron microscopy of these preparations revealed by far the most abundant and extensive arrays of intact collagen VI microfibrils isolated to date. The distinct microfibrillar species, fibrillin, which was identified on the basis of its periodicity and morphology, was also solubilized in abundance by this protocol. Analysis of these complex polymers has generated new information on their supramolecular architecture and relative abundance in these tissues. The protocol also demonstrates that the release of intact collagen VI microfibrils from these tissues is largely dependent on the removal of the major collagen fibrils.
Kielty
C M
CM
Department of Biochemistry and Molecular Biology, University of Manchester, UK.
Cummings
C
C
Whittaker
S P
SP
Shuttleworth
C A
CA
Grant
M E
ME
D000818:Animals; D001011:Aorta / Q000737:chemistry / Q000196:embryology; D002417:Cattle; D003094:Collagen / Q000737:chemistry / Q000302:isolation & purification* / Q000648:ultrastructure; D004547:Elastic Tissue / Q000737:chemistry / Q000196:embryology*; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000737:chemistry; D000071837:Fibrillins; D008022:Ligaments / Q000737:chemistry / Q000196:embryology; D046911:Macromolecular Substances; D008840:Microfilament Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008970:Molecular Weight; D012867:Skin / Q000737:chemistry / Q000196:embryology
D000071837:Fibrillins; D046911:Macromolecular Substances; D008840:Microfilament Proteins; D003094:Collagen