A unique family of cysteine proteases has been described that differs in sequence, structure and substrate specificity from any previously described protease family. This family, Ced-3/caspase-1, is composed of caspase-1, caspase-2, caspase-3, caspase-4, caspase-6 and caspase-7 (also designated Mch3, ICE-LAP3 or CMH-1), caspase-9, caspase-10, and caspase-14. Ced-3/caspase-1 family members function as key components of the apoptotic machinery and act to destroy specific target proteins which are critical to cellular longevity. Caspase-3, caspase-7 and caspase-9, but not caspase-1, have been shown to cleave the nuclear protein PARP into an apoptotic fragment. Caspase-14, also designated MICE (for mini-ICE), is highly expressed in embryonic tissues but appears to be absent from adult tissues. Procaspase-14 can be processed in vitro by caspase-8 and caspase-10 but not by other caspases.
Background References
1. Lemound, J. et al. 2015. Aberrant expression of caspase 14 in salivary gland carcinomas. J. Oral Pathol. Med.. 44: 444-8.
2. Pellerin, L. et al. 2014. Bleomycin hydrolase downregulation in lesional skin of adult atopic dermatitis patients is independent of FLG gene mutations. J. Allergy Clin. Immunol.. 134: 1459-1461.e7.
Sequence Similarity
Belongs to the peptidase C14A family.
Tissue Specificity
Expressed in keratinocytes of adult skin suprabasal layers (from spinous layers to the stratum granulosum and stratum corneum) (at protein level). Expressed in keratinocytes of hair shaft and sebaceous glands (at protein level). In psoriatic skin only expressed at very low levels. The p17/10 mature form is expressed in epidermis stratum corneum, the p20/p8 intermediate form in epidermis upper granular cells of the stratum granulosum.
Post-translational Modification
Maturation by proteolytic processing appears to be a two-step process. The precursor is processed by KLK7 to yield the p20/p8 intermediate form which acts on the precursor to yield the p17/p10 mature form. Initially, cleavage between Ile-152 and Lys-153 has been proposed to yield the large and small subunits of the active enzyme.
Western blot analysis of Caspase-14 on A431 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-52, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Western blot analysis of Caspase-14 on MCF-7 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-52, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
ICC staining of Caspase-14 in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-52, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ICC staining of Caspase-14 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-52, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ICC staining of Caspase-14 in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-52, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Caspase-14 antibody (ET1701-52) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-52) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Flow cytometric analysis of Caspase-14 was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1701-52, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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