Increased Ca2+ and myosin phosphorylation, but not calmodulin activity in sensitized airway smooth muscles.
The increased shortening velocity and capacity of airway smooth muscle (ASM) from ragweed pollen-sensitized dogs, which may be responsible for its in vivo airway hyperresponsiveness, have been shown to be associated with higher actomyosin adenosinetriphosphatase activity and greater level of phosphorylation of the 20-kDa myosin light-chain (MLC20) at rest and during contraction. Current studies show that the elevated level of phosphorylation may be the result of an increased myosin light-chain kinase (MLCK) activity due to excessive quantity of MLCK. There were no significant changes in total activity of calmodulin, a protein that binds and activates MLCK, in sensitized dog ASM (SASM) compared with control ASM (CASM). When normalized to the relative calmodulin content in the tissues, the specific calmodulin activities (means +/- SE) in sensitized tracheal smooth muscle (STSM) and sensitized bronchial smooth muscle (SBSM) and in their controls were not different (STSM 0.359 +/- 0.117, CTSM 0.339 +/- 0.136. SBSM 0.201 +/- 0.098, and control bronchial smooth muscle 0.213 +/- 0.056 nmol Pi.calmodulin content-1.min-1, respectively). Intracellular Ca2+ levels indicated by fura 2 fluorescent dye remained unaltered in SASM. We conclude that airway hyperresponsiveness may result from higher MLCK content in SASM rather than from changes in Ca(2+)-calmodulin activities, which is an example of alteration in Ca2- sensitivity of ASM.
Jiang
H
H
Department of Physiology, Faculty of Medicine, University of Manitoba, Winnipeg, Canada.
Rao
K
K
Liu
X
X
Liu
G
G
Stephens
N L
NL
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D004285:Dogs; D007114:Immunization; D007425:Intracellular Membranes / Q000378:metabolism; D009130:Muscle, Smooth / Q000276:immunology / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism; D009218:Myosins / Q000737:chemistry / Q000378:metabolism*; D009994:Osmolar Concentration; D010766:Phosphorylation; D013997:Time Factors; D014132:Trachea / Q000276:immunology / Q000378:metabolism*
D002147:Calmodulin; D009219:Myosin-Light-Chain Kinase; D009218:Myosins; D002118:Calcium