A new function of calphobindin I (annexin V). Promotion of both migration and urokinase-type plasminogen activator activity of normal human keratinocytes.
Calphobindin I (CPB-I, annexin V) is an anticoagulant protein purified from human placenta; it is a member of the annexin family that binds phospholipids in a calcium-dependent manner. In this study, we discovered and examined a new function of CPB-I: promotion of both the migration and the urokinase-type plasminogen activator (uPA) activity of normal human keratinocytes (NHK). While the treatment of NHK with a 10-micrograms/ml concentration of CPB-I for 24 h or 48 h caused an approximate 30% increase in the migration of NHK (compared with the no treatment), migration was inhibited when anti-CPB-I monoclonal antibodies (i.e. A46 and A180) were added along with the CPB-I. Moreover, while the treatment of NHK with a CPB-I concentration greater than 10 micrograms/ml caused a significant increase in the activity of secreted uPA, reflected in an approximately 40% increase in cell migration, uPA activity was inhibited both by cycloheximide and by monoclonal antibodies. This significant increase of secreted uPA was seen 8 h after the addition of CPB-I. Specific binding of CPB-I to NHK had a Kd value of 95.2 nM (equivalent to a CPB-I had no effect on NHK proliferation. Furthermore, CPB-I enhanced reepithelialization when it was applied locally twice a day to full-thickness cutaneous wounds made in male rats. Our results show that, during an injury, CPB-I helps reepithelialization through the promotion of both uPA synthesis and migration of keratinocytes without stimulating their proliferation.
Nakao
H
H
Department of Cell Biology, Kowa Research Institute, Ibaraki, Japan.
Watanabe
M
M
Maki
M
M
D000818:Animals; D017304:Annexin A5 / Q000378:metabolism / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects; D004848:Epithelium / Q000254:growth & development; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects*; D008297:Male; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D012867:Skin / Q000187:drug effects / Q000254:growth & development*; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis*; D014945:Wound Healing / Q000187:drug effects
D017304:Annexin A5; D014568:Urokinase-Type Plasminogen Activator