A role for zinc in regulating hypoxia-induced contractile events in pulmonary endothelium.

Abstract

We previously reported that zinc thiolate signaling contributes to hypoxic contraction of small, nonmuscularized arteries of the lung. The present studies were designed to investigate mechanisms by which hypoxia-released zinc induces contraction in isolated pulmonary endothelial cells and to delineate the signaling pathways involved in zinc-mediated changes in the actin cytoskeleton. We used fluorescence-based imaging to show that hypoxia induced time-dependent increases in actin stress fibers that were reversed by the zinc chelator, N,N,N',N'-tetrakis-(2-pyridylmethyl)-ethylenediamine (TPEN). We further showed that hypoxia-induced phosphorylation of the contractile protein myosin light chain (MLC) and assembly of actin stress fibers were each TPEN sensitive. Hypoxia and zinc-induced inhibition of MLC phosphatase (MLCP) were independent of the regulatory subunit (MYPT1) of MLCP, and therefore hypoxia-released zinc likely inhibits MLCP at its catalytic (PP1) subunit. Inhibition of PKC by Ro-31-8220 and a dominant-negative construct of PKC-ε attenuated hypoxia-induced contraction of isolated pulmonary endothelial cells. Furthermore, zinc-induced phosphorylation of MLC (secondary to inhibition of MLCP) was PKC dependent, and hypoxia-released zinc promoted the phosphorylation of the PKC substrate, CPI-17. Collectively, these data suggest a link between hypoxia, elevations in labile zinc, and activation of PKC, which in turn acts through CPI-17 to inhibit MLCP activity and promote MLC phosphorylation, ultimately inducing stress fiber formation and endothelial cell contraction.

• Authors: Paula J Bernal, Eileen M Bauer, Rong Cao, Salony Maniar, Mackenzie Mosher, Jun Chen, Qiming Jane Wang, Joseph C Glorioso, Bruce R Pitt, Simon C Watkins, Claudette M St Croix
• Journal: American journal of physiology. Lung cellular and molecular physiology (Am J Physiol Lung Cell Mol Physiol) Vol. 300 Issue 6 Pg. L874-86 (Jun 2011) ISSN: 1522-1504 [Electronic] United States
• PMID: 21378023 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Actins
• Muscle Proteins
• Phosphoproteins
• Ppp1r14a protein, rat
• Protein Kinase C
• Myosin-Light-Chain Kinase
• Myosin-Light-Chain Phosphatase
• Zinc

Topics

• Actins (metabolism)
• Animals
• Blotting, Western
• Cytoskeleton (metabolism)
• Endothelium, Vascular (cytology, drug effects, metabolism)
• Fluorescent Antibody Technique
• Hypoxia
• Muscle Contraction (drug effects)
• Muscle Proteins (metabolism)
• Myosin-Light-Chain Kinase (metabolism)
• Myosin-Light-Chain Phosphatase (metabolism)
• Phosphoproteins (metabolism)
• Protein Kinase C (metabolism)
• Pulmonary Artery (cytology, drug effects, metabolism)
• Rats
• Sheep
• Signal Transduction
• Stress Fibers
• Zinc (pharmacology)