Homo-oligomerization of human corneodesmosin is mediated by its N-terminal glycine loop domain.
Corneodesmosin (CDSN), a glycoprotein expressed during the late stages of epidermal differentiation, localizes in the extracellular core of upper desmosomes and of corneodesmosomes. Since it displays homophilic adhesive properties, CDSN is thought to reinforce cell-cell cohesion within the upper layers of the epidermis. CDSN presents two serine- and glycine-rich domains in its N- and C-terminus that may fold into highly flexible and adhesive secondary structures called glycine loops. We analyzed the importance of these domains in CDSN homophilic adhesion by producing full-length and truncated recombinant forms of the protein deleted of the N- and/or the C-terminal domain. The adhesive properties of the various proteins were then tested in vitro by overlay binding assays and surface plasmon resonance quantitative analysis. Experiments evidenced the homophilic adhesive properties of the N-terminal glycine loop domain, confirming its involvement in CDSN-CDSN interactions. They further indicated that most of the C-terminal domain is not necessary for the adhesive properties of the protein. The dissociation constant (K(D)) was calculated to be 1.3x10(-5) M. This interaction strength might allow dynamic regulation of the CDSN-CDSN association to occur in vivo. Moreover, molecular filtration analyses demonstrated for the first time that non-glycosylated CDSN is able to spontaneously form large homo-oligomers in vitro and that the N-terminal glycine loop domain is necessary for the formation of these macromolecular complexes.
Caubet
Cécile
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UMR 5165 Epidermis Differentiation and Rheumatoid Autoimmunity CNRS, University of Toulouse III, Institut Fédératif de Recherche Claude de Préval, IFR30 (INSERM-CNRS Université Paul Sabatier-Centre Hospitalier Universitaire), Toulouse, France.
Jonca
Nathalie
N
Lopez
Frédéric
F
Estève
Jean-Pierre
JP
Simon
Michel
M
Serre
Guy
G
D000595:Amino Acid Sequence; D019281:Dimerization; D005998:Glycine; D006023:Glycoproteins / Q000737:chemistry*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D020349:Surface Plasmon Resonance
C109656:CDSN protein, human; D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; D005998:Glycine