title: Type XVII collagen (BP180) and LAD-1 are present as separate trimeric complexes.
abstract: This study characterized the high molecular mass BP180 complex that is observed when unheated sodium dodecyl sulfate extracts of human skin or keratinocytes are subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. In heated extracts BP180 is present as a monomer with a molecular weight of 180 kDa, in unheated extracts BP180 runs at a molecular weight position over 500 kDa. By preincubating the unheated extracts at temperatures between 31 degrees C and 40 degrees C, the high molecular weight complex could be "melted" down to monomeric BP180. Under the conditions employed the T1/2 of the dissociation process was between 35 degrees C and 36 degrees C. The temperature resistance of the high molecular weight complex was used to analyze its molecular composition by performing two-dimensional electrophoresis with a "low-temperature" first dimension step and a "high-temperature" second dimension step. Silver staining and immunoblotting of the two-dimensional gels revealed the high molecular weight complex to be composed of solely BP180, indicating that the complex is the nondissociated homotrimeric form of BP180. The 120 kDa linear IgA dermatosis antigen (LAD-1) is an collagenous anchoring filament protein with homology to the extracellular collagenous part of BP180. Two-dimensional immunoblotting showed that LAD-1, as BP180, is also present as a high molecular mass complex and does not form mixed complexes with BP180.
authors.affiliation: Department of Dermatology, University Hospital, Groningen, The Netherlands.
authors.forename: H H
authors.initials: HH
authors.lastname: Pas
authors.forename: G J
authors.initials: GJ
authors.lastname: Kloosterhuis
authors.forename: M
authors.initials: M
authors.lastname: Nijenhuis
authors.forename: M C
authors.initials: MC
authors.lastname: de Jong
authors.forename: J B
authors.initials: JB
authors.lastname: van der Meer
authors.forename: M F
authors.initials: MF
authors.lastname: Jonkman
mesh_terms: D001324:Autoantigens / Q000737:chemistry*; D002352:Carrier Proteins; D003094:Collagen / Q000737:chemistry*; D003598:Cytoskeletal Proteins; D000071821:Dystonin; D006801:Humans; D008970:Molecular Weight; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D012872:Skin Diseases, Vesiculobullous / Q000276:immunology*; D013696:Temperature; D000097805:Collagen Type XVII
chemical_list: D001324:Autoantigens; D002352:Carrier Proteins; D003094:Collagen; D003598:Cytoskeletal Proteins; D000071821:Dystonin; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D000097805:Collagen Type XVII; C486268:DST protein, human