title: Caspase-14 protocols.
abstract: Unlike other caspase family members, caspase-14 shows restricted expression, being found mostly in epidermis and its appendages. It has been suggested that caspase-14 is not involved in apoptosis or inflammation, but participates in keratinocyte terminal differentiation. Its activation occurs at the corneocyte formation. In previous work, we have purified active caspase-14 from human corneocyte extracts. In addition, we have clarified activation mechanism of caspase-14, where kallikrein-related peptidase 7 (KLK7) generates an intermediate form from procaspase-14 and this form finally converts procaspase-14 to active, mature caspase-14. Here we describe techniques for measurement of caspase-14 activity using synthetic substrate, purification of caspase-14 from corneocyte extract, preparation of constitutively active caspase-14 and specific antibody, quantification of total and active caspase-14 in corneocyte extracts using ELISA, as well as methods for caspase-14 activation and its visualization by immunohistochemistry.
authors.affiliation: Shiseido Research Center, Tsuzuki-ku, Yokohama, Japan.
authors.forename: Mami
authors.initials: M
authors.lastname: Yamamoto-Tanaka
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Hibino
mesh_terms: D017209:Apoptosis / Q000235:genetics; D053456:Caspase 14 / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D004789:Enzyme Activation; D006801:Humans; D015603:Keratinocytes / Q000201:enzymology; D008967:Molecular Biology / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D011994:Recombinant Proteins; D053456:Caspase 14