Increased myofibrillar protein phosphatase-1 activity impairs rat aortic smooth muscle activation after hypoxia.
We hypothesized that increased myofibrillar type 1 protein phosphatase (PP1) catalytic activity contributes to impaired aortic smooth muscle contraction after hypoxia. Our results show that inhibition of PP1 activity with microcystin-LR (50 nmol/l) or okadaic acid (100 nmol/l) increased phenylephrine- and KCl-induced contraction to a greater extent in aortic rings from rats exposed to hypoxia (10% O(2)) for 48 h than in rings from normoxic animals. PP1 inhibition also restored the level of phosphorylation of the 20-kDa myosin light chain (LC(20)) during maximal phenylephrine-induced contraction to that observed in the normoxic control group. Myofibrillar PP1 activity was greater in aortas from rats exposed to hypoxia than in normoxic rats (P < 0.05). Levels of the protein myosin phosphatase-targeting subunit 1 (MYPT1) that mediates myofibrillar localization of PP1 activity were increased in aortas from hypoxic rats (193 +/- 28% of the normoxic control value, P < 0.05) and in human aortic smooth muscle cells after hypoxic (1% O(2)) incubation (182 +/- 18% of the normoxic control value, P < 0.05). Aortic levels of myosin light chain kinase were similar in normoxic and hypoxic groups. In conclusion, after hypoxia, increased MYPT1 protein and myofibrillar PP1 activity impair aortic vasoreactivity through enhanced dephosphorylation of LC(20).
Teoh
Hwee
H
Terrence Donnelly Laboratories, Division of Respirology and Department of Critical Care, St. Michael's Hospital, University of Toronto, Toronto, Ontario, Canada M5B 1W8.
Zacour
Mary
M
Wener
Avraham D
AD
Gunaratnam
Lakshman
L
Ward
Michael E
ME
D000818:Animals; D001011:Aorta; D015153:Blotting, Western; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D000860:Hypoxia / Q000503:physiopathology*; D008297:Male; D008387:Marine Toxins; D052998:Microcystins; D009119:Muscle Contraction / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000503:physiopathology*; D009210:Myofibrils / Q000201:enzymology*; D018994:Myosin Light Chains / Q000378:metabolism; D019319:Okadaic Acid / Q000494:pharmacology; D010100:Oxygen / Q000008:administration & dosage; D010456:Peptides, Cyclic / Q000494:pharmacology; D010656:Phenylephrine / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010766:Phosphorylation; D011189:Potassium Chloride / Q000494:pharmacology; D054645:Protein Phosphatase 1; D051381:Rats; D017207:Rats, Sprague-Dawley
D004791:Enzyme Inhibitors; D008387:Marine Toxins; D052998:Microcystins; D018994:Myosin Light Chains; D010456:Peptides, Cyclic; D019319:Okadaic Acid; D010656:Phenylephrine; D011189:Potassium Chloride; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1; C057862:cyanoginosin LR; D010100:Oxygen