SPINK9 stimulates metalloprotease/EGFR-dependent keratinocyte migration via purinergic receptor activation.
Serine protease inhibitors of the Kazal-type 9 (SPINK9) is a keratinocyte-derived cationic peptide that is found most abundantly in the upper layers of the palmar-plantar epidermis. In vitro, the peptide displays the capacity to inhibit specifically kallikrein-related peptidase 5 (KLK5). Here, we report that cells expressing SPINK9 secrete the peptide constitutively. Recombinant SPINK9 (rSPINK9) provoked transactivation of the EGFR in human keratinocytes, resulting in efficient downstream triggering of cell migration. Transactivation occurred via functional upregulation of a disintegrin and metalloproteases (ADAMs), as evidenced by suppression with a metalloproteinase inhibitor and an EGFR-blocking antibody. SPINK9 preparations isolated from human skin also displayed EGFR-transactivating capacity. The classical purinergic receptor antagonists oxidized ATP and pyridoxalphosphate-6-azophenyl-2',4',-disulfonic acid effectively suppressed EGFR transactivation by rSPINK9, indicating that in analogy to what has recently been reported for the cationic antimicrobial peptides cathelicidin LL-37 and bee venom melittin, purinergic receptors have an essential bridging role in promoting the upregulation of ADAM function by the cationic peptide. SPINK9 could represent an example of how a cationic peptide may subserve multiple and interrelated functions that contribute to the maintenance of the physical and immunological barrier of the skin.
Department of Dermatology, University Hospital Schleswig-Holstein, Christian-Albrecht University Kiel, Kiel, Germany.
Maria
M
Sperrhacke
Department of Dermatology, University Hospital Schleswig-Holstein, Christian-Albrecht University Kiel, Kiel, Germany.
Jan
J
Fischer
Department of Dermatology, University Hospital Schleswig-Holstein, Christian-Albrecht University Kiel, Kiel, Germany.
Zhihong
Z
Wu
Department of Dermatology, University Hospital Schleswig-Holstein, Christian-Albrecht University Kiel, Kiel, Germany.
Sarah
S
Klünder
Department of Transgenic Models of Diseases, Institute of Molecular Genetics of the ASCR, Prague, Czech Republic.
Radislav
R
Sedlacek
Department of Dermatology, University Hospital Schleswig-Holstein, Christian-Albrecht University Kiel, Kiel, Germany.
Jens-Michael
JM
Schroeder
Department of Dermatology, University Hospital Schleswig-Holstein, Christian-Albrecht University Kiel, Kiel, Germany.
Ulf
U
Meyer-Hoffert
Department of Dermatology, University Hospital Schleswig-Holstein, Christian-Albrecht University Kiel, Kiel, Germany. Electronic address: kreiss@dermatology.uni-kiel.de.
Karina
K
Reiss
D023181:Antimicrobial Cationic Peptides / Q000378:metabolism; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D003001:Cloning, Molecular; D066246:ErbB Receptors / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D057809:HEK293 Cells; D006801:Humans; D007610:Kallikreins / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology*; D045726:Metalloproteases / Q000378:metabolism; D053491:Proteinase Inhibitory Proteins, Secretory / Q000378:metabolism*; D011983:Receptors, Purinergic / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D000074162:Serine Peptidase Inhibitors, Kazal Type; D015398:Signal Transduction; D014162:Transfection; D014945:Wound Healing; D054804:Cathelicidins
D023181:Antimicrobial Cationic Peptides; D066246:ErbB Receptors; D007610:Kallikreins; D045726:Metalloproteases; D053491:Proteinase Inhibitory Proteins, Secretory; D011983:Receptors, Purinergic; D011994:Recombinant Proteins; D000074162:Serine Peptidase Inhibitors, Kazal Type; D054804:Cathelicidins; C542011:SPINK9 protein, human; C400147:KLK5 protein, human