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Vol. 20, No. 4, 2007
Issue release date: June 2007
Skin Pharmacol Appl Skin Physiol 2007;20:187–194

Kinetic Characteristics of Acidic and Alkaline Ceramidase in Human Epidermis

Houben E. · Uchida Y. · Nieuwenhuizen W.F. · De Paepe K. · Vanhaecke T. · Holleran W.M. · Rogiers V.
aDepartment of Toxicology, Dermato-Cosmetology and Pharmacognosy, Vrije Universiteit Brussel, Brussels, Belgium; bDepartment of Dermatology, University of California San Francisco and VA Medical Center, San Francisco, Calif., USA; cTNO Quality of Life, Zeist, The Netherlands

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It has recently become evident that at least five ceramidase (CDase) isoforms are present in human epidermis, and that specifically acidic CDase (aCDase) and alkaline CDase (alkCDase) activities increase during keratinocyte differentiation, and thus might play a pivotal role(s) in permeability barrier function. Prior to investigating their possible roles in the epidermal barrier function, it is necessary to characterize basic kinetic parameters for these enzymes, as well as to determine the effects of the established CDase inhibitors and their activities. In this study, assays for both aCDase and alkCDase activities in fully differentiated human epidermis were optimized using a radiolabeled substrate. These studies revealed that aCDase activity is substantially higher than alkCDase activity, and that both isoenzymes are inhibited by a CDase inhibitor N-oleylethanolamine. These findings were also confirmed using an in situ enzyme assay.

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