Purification and characterization of the endoglycosidase heparanase 1 from human plantar stratum corneum: a key enzyme in epidermal physiology?
A protein exhibiting endoglycosidase activity was purified from plantar stratum corneum to apparent homogeneity in two sequential column chromatographic steps. Protein sequencing revealed its identity with the recently cloned human heparanase 1, an enzyme, the expression of which is reported to be related to the metastasic potential of tumor cells. By using a heparanase 1 specific antibody we were able to demonstrate that, in the plantar stratum corneum, heparanase 1 exists in two forms, the active 50 kDa protein and the inactive 63 kDa form, probably a proform of the enzyme. The antibody also decorated numerous degradation fragments. Reverse transcription polymerase chain reaction studies as well as immunohistochemical analysis using reconstructed and normal human epidermis demonstrated clearly a keratinocyte differentiation related expression of heparanase 1. Interestingly, the antibody also strongly decorated dendritic cells, which after double labeling could be identified to be a subpopulation of the epidermal Langerhans cells. Based on our findings and the known history of this enzyme, we advanced the hypothesis that heparanase 1 has multiple physiologic functions in the epidermis: (i) it plays an important role in epidermal differentiation, possibly by modulating the liberation of heparan sulfate bound (growth) factors; (ii) in the stratum corneum, the endoglycosidase activity of heparanase 1 might be indispensable and represent the first step in the desquamation process; and (iii) in Langerhans cells, its catalytic activity is required for the trans-tissue migration of these cells.
Bernard
D
D
Life Sciences Research, L'Oréal, Center Charles Zviak, Clichy, France. dbernard@recherche.loreal.com
Méhul
B
B
Delattre
C
C
Simonetti
L
L
Thomas-Collignon
A
A
Schmidt
R
R
D000595:Amino Acid Sequence / Q000235:genetics; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology; D004817:Epidermis / Q000201:enzymology* / Q000502:physiology; D005528:Foot; D005966:Glucuronidase / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007527:Isoenzymes / Q000302:isolation & purification*; D015603:Keratinocytes / Q000166:cytology / Q000201:enzymology; D008969:Molecular Sequence Data; D014018:Tissue Distribution; D000099175:Heparanase
D005966:Glucuronidase; D007527:Isoenzymes; D000099175:Heparanase