The cornified envelope of the epidermal keratinocyte is a cross- linked protein structure which provides tensile strength to the stratum corneum and may be altered in some diseases. The research is this proposal will have two major goals. The foremost of these is to examine the hypothesis that the precursor proteins which make up the cornified envelope of the mature keratinocyte are similar across mammalian species and separable into classes based on charge, size, composition, and ability to be cross-linked by the enzyme transglutaminase. This hypothesis will be examined after fulfillment of the second major goal which is to identify, purify, and biochemically characterize the precursor proteins of murine cornified envelope as well as quantitate their contribution to the final product. The latter will be accomplished by generating monoclonal antibodies to whole cornified envelopes and using these antibodies as tools to identify precursors in the soluble and particulate fractions of murine keratinocyte preparations. The proteins will also be identified as transglutaminase substrates by their ability to cross-link with labeled aliphatic amines. Individual precursors will be isolated, purified and biochemically characterized. With this data, the former goal can be realized by direct comparison of biochemical properties between human, bovine, and murine precursors. Also, purified precursors from two species will be mixed with transglutaminase and the resultant reaction products examined for the presence of heteropolymers which would suggest that proteins from one species can functionally substitute for another in cross-linking reactions.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08AR001804-05
Application #
3079253
Study Section
Arthritis and Musculoskeletal and Skin Diseases Special Grants Review Committee (AMS)
Project Start
1988-09-01
Project End
1993-08-31
Budget Start
1992-09-01
Budget End
1993-08-31
Support Year
5
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02199
Greco, M A; Lorand, L; Lane, W S et al. (1995) The pancornulins: a group of small proline rich-related cornified envelope precursors with bifunctional capabilities in isopeptide bond formation. J Invest Dermatol 104:204-10
Ming, M E; Daryanani, H A; Roberts, L P et al. (1994) Binding of keratin intermediate filaments (K10) to the cornified envelope in mouse epidermis: implications for barrier function. J Invest Dermatol 103:780-4
Kvedar, J C; Manabe, M; Phillips, S B et al. (1992) Characterization of sciellin, a precursor to the cornified envelope of human keratinocytes. Differentiation 49:195-204
Baden, H P; Goldaber, M L; Kvedar, J C (1992) Keratinocytes stimulate prostaglandin I2 synthesis by 3T3 cells and exhibit enhanced cornification when exposed to prostaglandin I2 analogues. J Cell Physiol 150:269-75
Kvedar, J C; Pion, I A; Bilodeau, E B et al. (1992) Detection of substrates of keratinocyte transglutaminase in vitro and in vivo using a monoclonal antibody to dansylcadaverine. Biochemistry 31:49-56
Kvedar, J C; Daryanani, H A; Baden, H P (1991) Cross-linked components of the cuticle of the cortex of hair. J Invest Dermatol 96:84S-85S
Kvedar, J C; Baden, H P (1991) Nail changes in cutaneous disease. Semin Dermatol 10:65-70