Type VII collagen is a recently described collagen from human amnion. The biochemical and structural features of this molecule suggest that it is a component of anchoring fibrils. Anchoring fibrils are collagenase-sensitive, fibrous structures that are thought to anchor certain epithelial cell layers onto the underlying connective tissue. Anchoring fibrils are altered in many disease states, such as in recessive epidermoly sis bullosa dystrophica, a heritable skin disorder. This disorder is characterized by degeneration of the dermis and disruption of anchoring fibrils, resulting in severe blistering of the skin. Different regions of the dermal-epidermal junction are affected in patients with other forms of epidermolysis bullosa. The objectives of this research are (1) to define new proteins of the dermal-epidermal junction, (2) to identify the molecular components of the anchoring fibrils and (3) to determine the nature of the interaction between anchoring fibrils, the basement membrane and the underlying stroma.
Specific aims are (1) to isolate an anchoring fibril-basement membrane (AF-BM) complex from skin, using non-degradative, mechanical methods, (2) to solubilize, purify and characterize the various components of this complex, using standard techniques of self fractionation, column chromatography, HPLC and peptide mapping (3) to define the interactive sites on the molecules by performing binding studies using whole molecules and derived peptides and (4) to reconstitute anchoring fibrils from the purified components, monitoring fiber formation by electron microscopy. Human skin, fetal calf skin and skin from lathyritic chicken embryos will be used. Preliminary studies from this laboratory indicate that there is a collagenous protein in the AF-BM complex that resembles Type VII collagen. This protein will be compared to Type VII collagen from amnion to establish whether or not they are identical. In addition, there is a large, non-collagenous protein that differs from other previously described proteins of the epidermal-dermal junction. This protein will be characterized to define its chemical nature, precise location and ability to interact with other components of the AF-BM complex. Several monoclonal antibodies are available to aid in the localization of these proteins. These studies will help define the role of anchoring fibrils in the body and will provide a foundation for the study of connective tissue disorders that involve anchoring fibrils.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR035927-03
Application #
3157416
Study Section
Pathobiochemistry Study Section (PBC)
Project Start
1985-08-01
Project End
1989-01-31
Budget Start
1987-08-01
Budget End
1989-01-31
Support Year
3
Fiscal Year
1987
Total Cost
Indirect Cost
Name
University of California Los Angeles
Department
Type
Schools of Medicine
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Guthrie, C R; Murray, L W; Kopchok, G E et al. (1991) Biochemical mechanisms of laser vascular tissue fusion. J Invest Surg 4:3-12
Guthrie, C R; Murray, L W; Kopchok, G E et al. (1990) Alterations in biochemistry of laser-fused vascular tissue. Curr Surg 47:258-62
Glassberg, E; Lewandowski, L; Lask, G et al. (1990) Laser-induced photodynamic therapy with aluminum phthalocyanine tetrasulfonate as the photosensitizer: differential phototoxicity in normal and malignant human cells in vitro. J Invest Dermatol 94:604-10
Olsen, D R; Peltonen, J; Jaakkola, S et al. (1989) Collagen gene expression by cultured human skin fibroblasts. Abundant steady-state levels of type VI procollagen messenger RNAs. J Clin Invest 83:791-5
Murray, L W; Su, L; Kopchok, G E et al. (1989) Crosslinking of extracellular matrix proteins: a preliminary report on a possible mechanism of argon laser welding. Lasers Surg Med 9:490-6
Uitto, J; Olsen, D R; Fazio, M J (1989) Extracellular matrix of the skin: 50 years of progress. J Invest Dermatol 92:61S-77S
Uitto, J (1989) Connective tissue biochemistry of the aging dermis. Age-associated alterations in collagen and elastin. Clin Geriatr Med 5:127-47
Olsen, D; Nagayoshi, T; Fazio, M et al. (1989) Human laminin: cloning and sequence analysis of cDNAs encoding A, B1 and B2 chains, and expression of the corresponding genes in human skin and cultured cells. Lab Invest 60:772-82
Olsen, D R; Chu, M L; Uitto, J (1988) Expression of basement membrane zone genes coding for type IV procollagen and laminin by human skin fibroblasts in vitro: elevated alpha 1 (IV) collagen mRNA levels in lipoid proteinosis. J Invest Dermatol 90:734-8
Olsen, D R; Fazio, M J; Shamban, A T et al. (1988) Cutis laxa: reduced elastin gene expression in skin fibroblast cultures as determined by hybridizations with a homologous cDNA and an exon 1-specific oligonucleotide. J Biol Chem 263:6465-7

Showing the most recent 10 out of 11 publications