The anchoring complex is an ultrastructural feature of the dermal-epidermal basement membrane that includes the hemidesmosomes, anchoring filaments and anchoring fibrils. These structures are believed to work as a unit to stabilize epithelial attachment to the underlying basement membrane. Failure of components of the anchoring complex results in spontaneous blistering characteristic of the inherited bullous diseases. We have recently identified two laminin variants that are components of the anchoring filaments. We have termed these kalinin and K-laminin. We know that kalinin is absent from the basement membrane zone of patients with lethal epidermolysis bullosa. Kalinin is a cell adhesion macromolecule specific for certain types of epithelial cells including keratinocytes. For these cells, kalinin substitutes for laminin as the favored growth substrate. Monoclonal antibodies made to kalinin cause cell detachment and de-epithelialization of whole skin in vitro. This application is focused upon continued characterization of the structure and function of kalinin and K-laminin. We propose to isolate kalinin and K-laminin for protein sequence determinations and for preparing protein fragments; to complete the deduced amino acid sequences of these chains and to express selected subdomains in vitro; and to generate domain specific monoclonal antibodies. These reagents will be used to evaluate the binding function of kalinin and K-laminin subdomains. We will interfere with these functions in an in vitro skin equivalent model. These probes will also be used to evaluate mutations in the kalinin and K-laminin chains relative to the pathogenesis of junctional epidermolysis bullosa.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37AR035689-14
Application #
2542941
Study Section
Special Emphasis Panel (NSS)
Project Start
1985-09-23
Project End
2003-04-30
Budget Start
1998-05-01
Budget End
1999-04-30
Support Year
14
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02199
Koch, M; Murrell, J R; Hunter, D D et al. (2000) A novel member of the netrin family, beta-netrin, shares homology with the beta chain of laminin: identification, expression, and functional characterization. J Cell Biol 151:221-34
Amano, S; Scott, I C; Takahara, K et al. (2000) Bone morphogenetic protein 1 is an extracellular processing enzyme of the laminin 5 gamma 2 chain. J Biol Chem 275:22728-35
Koch, M; Olson, P F; Albus, A et al. (1999) Characterization and expression of the laminin gamma3 chain: a novel, non-basement membrane-associated, laminin chain. J Cell Biol 145:605-18
Champliaud, M F; Burgeson, R E; Jin, W et al. (1998) cDNA cloning and characterization of sciellin, a LIM domain protein of the keratinocyte cornified envelope. J Biol Chem 273:31547-54
Cserhalmi-Friedman, P B; Baden, H; Burgeson, R E et al. (1998) Molecular basis of non-lethal junctional epidermolysis bullosa: identification of a 38 basepair insertion and a splice site mutation in exon 14 of the LAMB3 gene. Exp Dermatol 7:105-11
Wewer, U M; Thornell, L E; Loechel, F et al. (1997) Extrasynaptic location of laminin beta 2 chain in developing and adult human skeletal muscle. Am J Pathol 151:621-31
Gerecke, D R; Olson, P F; Koch, M et al. (1997) Complete primary structure of two splice variants of collagen XII, and assignment of alpha 1(XII) collagen (COL12A1), alpha 1(IX) collagen (COL9A1), and alpha 1(XIX) collagen (COL19A1) to human chromosome 6q12-q13. Genomics 41:236-42
Rousselle, P; Keene, D R; Ruggiero, F et al. (1997) Laminin 5 binds the NC-1 domain of type VII collagen. J Cell Biol 138:719-28
Champliaud, M F; Lunstrum, G P; Rousselle, P et al. (1996) Human amnion contains a novel laminin variant, laminin 7, which like laminin 6, covalently associates with laminin 5 to promote stable epithelial-stromal attachment. J Cell Biol 132:1189-98
Marinkovich, M P; Taylor, T B; Keene, D R et al. (1996) LAD-1, the linear IgA bullous dermatosis autoantigen, is a novel 120-kDa anchoring filament protein synthesized by epidermal cells. J Invest Dermatol 106:734-8

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