High-affinity binding of the NC1 domain of collagen VII to laminin 5 and collagen IV
Section snippets
Materials and methods
Proteins for the binding studies. Recombinant mini-procollagen VII (mProVII) consisting of the NC1 domain, a truncated triple-helical region, and the NC2 domain was purified from cell culture media as described [13]. To express recombinant NC1 domain of human collagen VII, we employed a DNA fragment encompassing a 1–3870 bp2 region of cDNA encoding human procollagen VII (a kind gift from Dr. Y. Gache, Universite de
Proteins for biosensor binding assays
As demonstrated in Fig. 1, the NC1 domain had a predicted molecular mass of 137 kDa [12]. Electrophoresis in nonreducing conditions revealed that some of the NC1 molecules formed homotypic complexes stabilized by the disulfide bonds (Fig. 1) formed between the cysteine residues present within the NC1 domain [12]. Electron microscopy of the recombinant NC1 domains (Fig. 2) confirmed that they had a structure similar to that of the native NC1 domains seen in mProVII. Specifically, the NC1 domains
Discussion
Noncovalent interactions between macromolecules are essential for the formation of supramolecular structures of biological scaffolds that shape the architecture of tissues and organs. A macromolecular scaffold of skin is a complex structure formed by a number of molecules interacting with each other in a site-specific manner. One of the distinct morphological features of skin is that it is composed of two principal layers, namely the epidermis and dermis, separated by the basement membrane, a
Acknowledgments
We thank Dr. Yannick Gache for DNA constructs encoding the NC1 domain, Dr. David E. Birk and Biao Zuo for expert technical assistance with the electron microscopy. This study was supported by the NIH/NIAMS Grant P01-AR38923.
References (36)
- et al.
The dermal–epidermal junction
Curr. Opin. Cell Biol.
(1997) - et al.
Laminins of the dermo–epidermal junction
Matrix Biol.
(1999) - et al.
Basement-membrane stromal relationships: interactions between collagen fibrils and the lamina densa
Int. Rev. Cytol.
(1997) - et al.
Molecular organization of the cutaneous basement membrane zone
Clin. Dermatol.
(2001) - et al.
Epiligrin, a new cell adhesion ligand for integrin alpha 3 beta 1 in epithelial basement membranes
Cell
(1991) - et al.
Nidogen mediates the formation of ternary complexes of basement membrane components
Kidney Int.
(1993) - et al.
Cloning of human type VII collagen. Complete primary sequence of the alpha 1(VII) chain and identification of intragenic polymorphisms
J. Biol. Chem.
(1994) - et al.
Proteinases of the bone morphogenetic protein-1 family convert procollagen VII to mature anchoring fibril collagen
J. Biol. Chem.
(2002) - et al.
NC1 domain of type VII collagen binds to the beta3 chain of laminin 5 via a unique subdomain within the fibronectin-like repeats
J. Invest. Dermatol.
(1999) - et al.
Interactions of the amino-terminal noncollagenous (NC1) domain of type VII collagen with extracellular matrix components. A potential role in epidermal–dermal adherence in human skin
J. Biol. Chem.
(1997)
Progress in heritable skin diseases: molecular bases and clinical implications
J. Am. Acad. Dermatol.
Single amino acid substitutions in procollagen VII affect early stages of assembly of anchoring fibrils
J. Biol. Chem.
Evidence that anti-type VII collagen antibodies are pathogenic and responsible for the clinical, histological, and immunological features of epidermolysis bullosa acquisita
J. Invest. Dermatol.
Structural determinants of the selectivity of KTS-disintegrins for the alpha1beta1 integrin
FEBS Lett.
Copolymerization of normal type I collagen with three mutated type I collagens containing substitutions of cysteine at different glycine positions in the alpha 1 (I) chain
J. Biol. Chem.
CLAMP: a biosensor kinetic data analysis program
Trends Biochem. Sci.
A substitution of cysteine for glycine 748 of the alpha 1 chain produces a kink at this site in the procollagen I molecule and an altered N-proteinase cleavage site over 225 nm away
J. Biol. Chem.
The epidermal–dermal junction
J. Invest. Dermatol.
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