title: Connective tissue microfibrils. Isolation and characterization of three large pepsin-resistant domains of fibrillin.
abstract: Human amnion was solubilized using pepsin and the digest supernatant screened for fragments of fibrillin with a previously characterized monoclonal antibody (Sakai, L. Y., Keene, D. R., and Engvall, E. (1986) J. Cell Biol. 103, 2499-2509). One fragment (PF1), with an apparent molecular weight of 94,000, was isolated and characterized. Two other fragments, PF2 and PF3, were isolated and shown to be fragments of fibrillin by preparing a monospecific antisera to PF2 and a monoclonal antibody to PF3. Immunoelectron microscopy and immunoblotting showed that both antibodies were specific for fibrillin. Electron microscope pictures of rotary-shadowed PF1 and PF2 showed them to be short rod-shaped molecules while PF3 has a crab-like appearance and seems to be an aggregate of several fibrillin chain fragments. Amino-terminal amino acid sequencing of PF1 and PF2 gave single unique sequences. Each of the three antibodies used was specific for one fragment and peptide mapping of PF1 and PF2 showed that there was no significant amino acid sequence overlap. Aggregates of PF3 are described which provided insight into the assembly and macromolecular structure of fibrillin in microfibrils.
authors.affiliation: Shriners Hospital for Crippled Children, Portland, Oregon 97201.
authors.forename: B K
authors.initials: BK
authors.lastname: Maddox
authors.forename: L Y
authors.initials: LY
authors.lastname: Sakai
authors.forename: D R
authors.initials: DR
authors.lastname: Keene
authors.forename: R W
authors.initials: RW
authors.lastname: Glanville
mesh_terms: D008841:Actin Cytoskeleton / Q000648:ultrastructure; D000596:Amino Acids / Q000032:analysis; D000650:Amnion / Q000032:analysis / Q000648:ultrastructure; D005260:Female; D000071837:Fibrillins; D006801:Humans; D008840:Microfilament Proteins / Q000302:isolation & purification / Q000648:ultrastructure; D008854:Microscopy, Electron; D010434:Pepsin A / Q000378:metabolism; D010449:Peptide Mapping; D011247:Pregnancy; D012867:Skin / Q000648:ultrastructure; D013379:Substrate Specificity
chemical_list: D000596:Amino Acids; D000071837:Fibrillins; D008840:Microfilament Proteins; D010434:Pepsin A