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Identification of lympho-epithelial Kazal-type inhibitor 2 in human skin as a kallikrein-related peptidase 5-specific protease inhibitor.

Meyer-Hoffert U, Wu Z, Schröder JM - PLoS ONE (2009)

Bottom Line: Recombinant LEKTI-2 inhibited KLK5 but not KLK7, 14 or other serine proteases tested including trypsin, plasmin and thrombin.LEKTI-2 immune-expression was focally localized at the stratum granulosum and stratum corneum at palmar and plantar sites in close localization to KLK5.At sites of plantar hyperkeratosis, LEKTI-2 expression was increased.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany.

ABSTRACT
Kallikreins-related peptidases (KLKs) are serine proteases and have been implicated in the desquamation process of the skin. Their activity is tightly controlled by epidermal protease inhibitors like the lympho-epithelial Kazal-type inhibitor (LEKTI). Defects of the LEKTI-encoding gene serine protease inhibitor Kazal type (Spink)5 lead to the absence of LEKTI and result in the genodermatose Netherton syndrome, which mimics the common skin disease atopic dermatitis. Since many KLKs are expressed in human skin with KLK5 being considered as one of the most important KLKs in skin desquamation, we proposed that more inhibitors are present in human skin. Herein, we purified from human stratum corneum by HPLC techniques a new KLK5-inhibiting peptide encoded by a member of the Spink family, designated as Spink9 located on chromosome 5p33.1. This peptide is highly homologous to LEKTI and was termed LEKTI-2. Recombinant LEKTI-2 inhibited KLK5 but not KLK7, 14 or other serine proteases tested including trypsin, plasmin and thrombin. Spink9 mRNA expression was detected in human skin samples and in cultured keratinocytes. LEKTI-2 immune-expression was focally localized at the stratum granulosum and stratum corneum at palmar and plantar sites in close localization to KLK5. At sites of plantar hyperkeratosis, LEKTI-2 expression was increased. We suggest that LEKTI-2 contributes to the regulation of the desquamation process in human skin by specifically inhibiting KLK5.

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LEKTI-2 is expressed at palmar and plantar sites.Immunohistochemical staining (Vector red) of LEKTI-2 of paraffin-embedded human skin samples using polyclonal antibodies. Only palmar and plantar localizations (A, n = 8) exhibited obvious LEKTI-2 immunoreactivity at the stratum granulosum and stratum corneum. Other localizations (e.g. trunk, B, n = 16) did not reveal LEKTI-2 immunoreactivity. Bars indicate 50 µm.
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pone-0004372-g005: LEKTI-2 is expressed at palmar and plantar sites.Immunohistochemical staining (Vector red) of LEKTI-2 of paraffin-embedded human skin samples using polyclonal antibodies. Only palmar and plantar localizations (A, n = 8) exhibited obvious LEKTI-2 immunoreactivity at the stratum granulosum and stratum corneum. Other localizations (e.g. trunk, B, n = 16) did not reveal LEKTI-2 immunoreactivity. Bars indicate 50 µm.

Mentions: To analyze LEKTI-2 protein expression, we generated affinity-purified polyclonal LEKTI-2 antibodies. Westernblot analyses performed with rLEKTI-2, purified natural LEKTI-2 and stratum corneum extracts revealed antigen specificity of the antibodies (Fig. 4), which was further confirmed by blocking experiments using recombinant LEKTI-2. Subsequently, LEKTI-2 immunohistochemistry was performed to localize LEKTI-2 expression in human skin samples. LEKTI-2 immunoreactivity was detected in the stratum granulosum and SC of human skin at the palms (inner sides) of hands and feet (n = 8) but no visible immunoreactivity was detected at other sites of healthy human skin (n = 16) (Fig. 5).


Identification of lympho-epithelial Kazal-type inhibitor 2 in human skin as a kallikrein-related peptidase 5-specific protease inhibitor.

Meyer-Hoffert U, Wu Z, Schröder JM - PLoS ONE (2009)

LEKTI-2 is expressed at palmar and plantar sites.Immunohistochemical staining (Vector red) of LEKTI-2 of paraffin-embedded human skin samples using polyclonal antibodies. Only palmar and plantar localizations (A, n = 8) exhibited obvious LEKTI-2 immunoreactivity at the stratum granulosum and stratum corneum. Other localizations (e.g. trunk, B, n = 16) did not reveal LEKTI-2 immunoreactivity. Bars indicate 50 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2631147&req=5

pone-0004372-g005: LEKTI-2 is expressed at palmar and plantar sites.Immunohistochemical staining (Vector red) of LEKTI-2 of paraffin-embedded human skin samples using polyclonal antibodies. Only palmar and plantar localizations (A, n = 8) exhibited obvious LEKTI-2 immunoreactivity at the stratum granulosum and stratum corneum. Other localizations (e.g. trunk, B, n = 16) did not reveal LEKTI-2 immunoreactivity. Bars indicate 50 µm.
Mentions: To analyze LEKTI-2 protein expression, we generated affinity-purified polyclonal LEKTI-2 antibodies. Westernblot analyses performed with rLEKTI-2, purified natural LEKTI-2 and stratum corneum extracts revealed antigen specificity of the antibodies (Fig. 4), which was further confirmed by blocking experiments using recombinant LEKTI-2. Subsequently, LEKTI-2 immunohistochemistry was performed to localize LEKTI-2 expression in human skin samples. LEKTI-2 immunoreactivity was detected in the stratum granulosum and SC of human skin at the palms (inner sides) of hands and feet (n = 8) but no visible immunoreactivity was detected at other sites of healthy human skin (n = 16) (Fig. 5).

Bottom Line: Recombinant LEKTI-2 inhibited KLK5 but not KLK7, 14 or other serine proteases tested including trypsin, plasmin and thrombin.LEKTI-2 immune-expression was focally localized at the stratum granulosum and stratum corneum at palmar and plantar sites in close localization to KLK5.At sites of plantar hyperkeratosis, LEKTI-2 expression was increased.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany.

ABSTRACT
Kallikreins-related peptidases (KLKs) are serine proteases and have been implicated in the desquamation process of the skin. Their activity is tightly controlled by epidermal protease inhibitors like the lympho-epithelial Kazal-type inhibitor (LEKTI). Defects of the LEKTI-encoding gene serine protease inhibitor Kazal type (Spink)5 lead to the absence of LEKTI and result in the genodermatose Netherton syndrome, which mimics the common skin disease atopic dermatitis. Since many KLKs are expressed in human skin with KLK5 being considered as one of the most important KLKs in skin desquamation, we proposed that more inhibitors are present in human skin. Herein, we purified from human stratum corneum by HPLC techniques a new KLK5-inhibiting peptide encoded by a member of the Spink family, designated as Spink9 located on chromosome 5p33.1. This peptide is highly homologous to LEKTI and was termed LEKTI-2. Recombinant LEKTI-2 inhibited KLK5 but not KLK7, 14 or other serine proteases tested including trypsin, plasmin and thrombin. Spink9 mRNA expression was detected in human skin samples and in cultured keratinocytes. LEKTI-2 immune-expression was focally localized at the stratum granulosum and stratum corneum at palmar and plantar sites in close localization to KLK5. At sites of plantar hyperkeratosis, LEKTI-2 expression was increased. We suggest that LEKTI-2 contributes to the regulation of the desquamation process in human skin by specifically inhibiting KLK5.

Show MeSH
Related in: MedlinePlus