CAP37 activation of PKC promotes human corneal epithelial cell chemotaxis.

PURPOSE:The objective of this study was to elucidate the signaling pathway through which cationic antimicrobial protein of 37 kDa (CAP37) mediates human corneal epithelial cell (HCEC) chemotaxis. METHODS:Immortalized HCECs were treated with pertussis toxin (10 and 1000 ng/mL), protein kinase C inhibitors (calphostin c, 50 nM and Ro-31-8220, 100 nM), phorbol esters (phorbol 12,13-dibutyrate, 200 nM and phorbol 12-myristate 13-acetate, 1 μM) known to deplete isoforms, and siRNAs (400 nM) before a modified Boyden chamber assay was used to determine the effect of these inhibitors and siRNAs on CAP37-directed HCEC migration. protein levels, phosphorylation, and duanyu1531δ kinase activity was assessed in CAP37-treated HCECs using immunohistochemistry, Western blotting, and a kinase activity assay, respectively. RESULTS:Chemotaxis studies revealed that treatment with pertussis toxin, duanyu1531 inhibitors, phorbol esters, and siRNAs significantly inhibited CAP37-mediated chemotaxis compared with untreated controls. CAP37 treatment increased duanyu1531δ protein levels and led to duanyu1531δ phosphorylation on residue Thr(505). Direct activation of duanyu1531δ by CAP37 was demonstrated using a kinase activity assay. CONCLUSIONS:These findings lead us to conclude that CAP37 is an important regulator of corneal epithelial cell migration and mediates its effects through

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