Calpain regulates neutrophil chemotaxis.
Cell polarization is required for directed cell migration. We investigated the role of the calcium-dependent protease calpain during neutrophil chemotaxis and found that calpain inhibition induced neutrophil adhesion, polarization, and rapid chemokinesis in the absence of exogenous activators. Resting neutrophils display constitutive calpain activity with mu-calpain being the predominant active isoform. Our findings suggest that constitutive calpain activity in resting neutrophils may function as a negative regulator of protrusion and migration. Specific inhibition of mu-calpain, but not m-calpain, induced neutrophil polarization and chemokinesis. In contrast to IL-8-induced chemokinesis, the chemokinesis induced by calpain inhibition was not reduced in the presence of pertussis toxin, suggesting that calpain functions downstream of G protein-coupled receptors. Further, both calpain inhibition and stimulation with IL-8 and formyl-Met-Leu-Phe (fMLP) induced an increase in Cdc42 and Rac activation. These findings are consistent with the involvement of calpain in chemotaxis pathways. Accordingly, calpain inhibition decreased neutrophil chemotaxis and directional persistence in a gradient of IL-8 and fMLP. Together, these data reveal a previously uncharacterized function for calpain in neutrophils and suggest that localized modulation of calpain activity may regulate neutrophil chemotaxis downstream of G-protein-coupled receptors.
Lokuta
M A
MA
Department of Pediatrics, University of Wisconsin, 1300 University Avenue, Madison, WI 53706, USA. malokuta@facstaff.wisc.edu
Nuzzi
P A
PA
Huttenlocher
A
A
D002154:Calpain / Q000097:blood*; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D016764:Cell Polarity; D002634:Chemotaxis, Leukocyte / Q000187:drug effects / Q000502:physiology*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D007930:Leucine / Q000031:analogs & derivatives* / Q000494:pharmacology; D018715:Microscopy, Video; D009504:Neutrophils / Q000166:cytology / Q000502:physiology*; D011480:Protease Inhibitors / Q000494:pharmacology
D015853:Cysteine Proteinase Inhibitors; D011480:Protease Inhibitors; C116501:acetyl-leucinal-leucinal-normethional; D002154:Calpain; C449000:m-calpain; C449002:mu-calpain; D007930:Leucine; D004121:Dimethyl Sulfoxide