Telokin expression and the effect of hypoxia on its phosphorylation status in smooth muscle cells from small and large pulmonary arteries.
Small pulmonary arteries (SPA), <500 microm diameter of the cat, constrict when exposed to hypoxia, whereas larger arteries (large pulmonary arteries; LPA), >800 microm diameter, show little or no response. It is unknown why different contractile responses occur within the same vascular bed, but activator or repressor proteins within the smooth muscle cell (SMC) can modify myosin phosphatase and myosin light chain kinase (MLCK), thereby influencing the phosphorylation state of myosin light chain (MLC) and ultimately, contraction. Telokin, a protein with a sequence identical to the COOH-terminal domain of MLCK, is expressed in smooth muscle where in its phosphorylated state it inhibits myosin phosphatase, binds to unphosphorylated myosin, and helps maintain smooth muscle relaxation. We measured telokin mRNA and telokin protein in smooth muscle from different diameter feline pulmonary arteries and sought to determine whether changes in the phosphorylation status of telokin and MLC occurred during hypoxia. In pulmonary arteries, telokin expression varied inversely with artery diameter, but cerebral arteries showed neither telokin protein nor telokin mRNA. Although telokin and MLC were distributed uniformly throughout the SPA muscle cell cytoplasm, they were not colocalized. During hypoxia, telokin dephosphorylated, and MLC became increasingly phosphorylated in SPA SMC, whereas in LPA SMC there was no change in either telokin or MLC phosphorylation. When LPA SMC were exposed to phenylephrine, MLC phosphorylation increased with no change in telokin phosphorylation. These results suggest that in SPA, phosphorylated telokin may help maintain relaxation under unstimulated conditions, whereas in LPA, telokin's function remains undetermined.
Madden
Jane A
JA
Department of Neurology, Zablocki Veterans Affairs Medical Center, Milwaukee, Wisconsin 53295, USA. jmadden@mcw.edu
Dantuma
Mark W
MW
Sorokina
Elena A
EA
Weihrauch
Dorothee
D
Kleinman
Jack G
JG
D000818:Animals; D002415:Cats; D002478:Cells, Cultured; D002536:Cerebral Arteries / Q000166:cytology / Q000502:physiology; D005260:Female; D015870:Gene Expression; D000860:Hypoxia / Q000503:physiopathology*; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism; D009219:Myosin-Light-Chain Kinase / Q000096:biosynthesis* / Q000378:metabolism; D010446:Peptide Fragments / Q000096:biosynthesis* / Q000378:metabolism; D010766:Phosphorylation; D011651:Pulmonary Artery / Q000166:cytology / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism
D010446:Peptide Fragments; D012333:RNA, Messenger; C060640:telokin; D009219:Myosin-Light-Chain Kinase