Organization of the transcortin-binding domain on placental plasma membranes.
Complex formation between transcortin (corticosteroid-binding globulin) and 20 kDa sialoglycoprotein from human syncytiotrophoblast plasma membranes (presumably a transcortin-recognizing subunit of the transcortin membrane receptor) was studied using FPLC and cross-linking with bifunctional reagents. The action of 1,5-difluoro-2,4-dinitrobenzene (DFDNB) on a solution of the purified 20 kDa sialoglycoprotein and transcortin resulted in formation of covalently linked complexes of 95 kDa and 140 kDa consisting of one transcortin molecule and either two or four molecules of the membrane sialoglycoprotein (the molecular mass of transcortin is 55 kDa). Additionally, cross-linking resulted in the appearance of a 43 kDa species which is the cross-linked dimer of the membrane protein. The dimer was also observed during chromatography on a Superose 12 column in the absence of DFDNB treatment. Treatment of intact syncytiotrophoblast membranes with DFDNB resulted in isolation of the transcortin binding protein dimer as the major portion of total pool of the protein. Formation of the transcortin complexes with two and four molecules of the membrane protein was also observed when the membranes were incubated with 125I-labeled transcortin and treated with DFDNB, but formation of the latter complexes predominated. The results obtained suggest that the recognizing and binding domain for transcortin in placental membranes is organized as dimers consisting of non-covalently linked sialoglycoprotein monomers of a 20 kDa each and that transcortin has two sites for interaction with this dimer. Apparently, binding of two dimers results in the formation of the functional form of the transcortin-receptor complex. The possible biological role of such a complex is discussed.
Krupenko
S A
SA
Institute of Biorganic Chemistry, Byelorussian Academy of Sciences, Minsk, Belarus.
Kolesnik
O I
OI
Krupenko
N I
NI
Strel'chyonok
O A
OA
D001665:Binding Sites; D002462:Cell Membrane / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents; D004120:Dimethyl Suberimidate; D004139:Dinitrofluorobenzene; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D046911:Macromolecular Substances; D008970:Molecular Weight; D010920:Placenta / Q000648:ultrastructure*; D011247:Pregnancy; D011956:Receptors, Cell Surface / Q000737:chemistry*; D015843:Serpins; D014156:Transcortin
D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D011956:Receptors, Cell Surface; C534935:SERPINA6 protein, human; D015843:Serpins; D004120:Dimethyl Suberimidate; D014156:Transcortin; D004139:Dinitrofluorobenzene