An increase or a decrease in myosin II phosphorylation inhibits macrophage motility.
Myosin II purified from mammalian non-muscle cells is phosphorylated on the 20-kD light chain subunit (MLC20) by the Ca2+/calmodulin-dependent enzyme myosin light chain kinase (MLCK). The importance of MLC20 phosphorylation in regulating cell motility was investigated by introducing either antibodies to MLCK (MK-Ab) or a Ca2+/calmodulin-independent, constitutively active form of MLCK (MK-) into macrophages. The effects of these proteins on cell motility were then determined using a quantitative chemotaxis assay. Chemotaxis is significantly diminished in macrophages containing MK-Ab compared to macrophages containing control antibodies. Moreover, there is an inverse relationship between the number of cells that migrate and the amount of MK-Ab introduced into cells. Interestingly, there is also an inverse relationship between the number of cells that migrate and the amount of MK- introduced into cells. Other experiments demonstrated that MK-Ab decreased intracellular MLC20 phosphorylation while MK- increased MLC20 phosphorylation. MK- also increased the amount of myosin associated with the cytoskeleton. These data demonstrate that the regulation of MLCK is an important aspect of cell motility and suggest that MLC20 phosphorylation must be maintained within narrow limits during translational motility by mammalian cells.
Wilson
A K
AK
Department of Physiology, College of Medicine, University of Illinois, Chicago 60680.
Gorgas
G
G
Claypool
W D
WD
de Lanerolle
P
P
D000818:Animals; D000906:Antibodies / Q000502:physiology; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D002633:Chemotaxis / Q000187:drug effects / Q000502:physiology; D008264:Macrophages / Q000187:drug effects / Q000502:physiology*; D008297:Male; D009219:Myosin-Light-Chain Kinase / Q000276:immunology / Q000502:physiology; D009218:Myosins / Q000378:metabolism* / Q000502:physiology; D010766:Phosphorylation; D051381:Rats; D011919:Rats, Inbred Strains
D000906:Antibodies; D009219:Myosin-Light-Chain Kinase; D009218:Myosins