Barrier-Protective Effects of Activated Protein C in Human Alveolar Epithelial Cells

Puig, Ferranda; Fuster, Gemma; Adda, Mélanie; Blanch, Lluís; Farre, Ramon; Navajas, Daniel; Artigas, Antonio
February 2013
PLoS ONE;Feb2013, Vol. 8 Issue 2, p1
Academic Journal
Acute lung injury (ALI) is a clinical manifestation of respiratory failure, caused by lung inflammation and the disruption of the alveolar-capillary barrier. Preservation of the physical integrity of the alveolar epithelial monolayer is of critical importance to prevent alveolar edema. Barrier integrity depends largely on the balance between physical forces on cell-cell and cell-matrix contacts, and this balance might be affected by alterations in the coagulation cascade in patients with ALI. We aimed to study the effects of activated protein C (APC) on mechanical tension and barrier integrity in human alveolar epithelial cells (A549) exposed to thrombin. Cells were pretreated for 3 h with APC (50 µg/ml) or vehicle (control). Subsequently, thrombin (50 nM) or medium was added to the cell culture. APC significantly reduced thrombin-induced cell monolayer permeability, cell stiffening, and cell contraction, measured by electrical impedance, optical magnetic twisting cytometry, and traction microscopy, respectively, suggesting a barrier-protective response. The dynamics of the barrier integrity was also assessed by western blotting and immunofluorescence analysis of the tight junction ZO-1. Thrombin resulted in more elongated ZO-1 aggregates at cell-cell interface areas and induced an increase in ZO-1 membrane protein content. APC attenuated the length of these ZO-1 aggregates and reduced the ZO-1 membrane protein levels induced by thrombin. In conclusion, pretreatment with APC reduced the disruption of barrier integrity induced by thrombin, thus contributing to alveolar epithelial barrier protection.


Related Articles

  • Theoretical study of structural changes caused by applying mechanical strain on peptide L. Krajčí, Miroslav; Urban, Ján; Mach, Pavel // Journal of Molecular Modeling;Nov2010, Vol. 16 Issue 11, p1693 

    The influence of the mechanical strain on the artificial protein L (acetyl-K-L-K-amide) has been studied at the molecular mechanics (MM) level of theory. The effect of the surrounding environment (DPPC molecules) has been observed during the stretching or compressing of the L. The calculations...

  • The mammalian-membrane two-hybrid assay (MaMTH) for probing membrane-protein interactions in human cells. Petschnigg, Julia; Groisman, Bella; Kotlyar, Max; Taipale, Mikko; Zheng, Yong; Kurat, Christoph F; Sayad, Azin; Sierra, J Rafael; Usaj, Mojca Mattiazzi; Snider, Jamie; Nachman, Alex; Krykbaeva, Irina; Tsao, Ming-Sound; Moffat, Jason; Pawson, Tony; Lindquist, Susan; Jurisica, Igor; Stagljar, Igor // Nature Methods;May2014, Vol. 11 Issue 5, p585 

    Cell signaling, one of key processes in both normal cellular function and disease, is coordinated by numerous interactions between membrane proteins that change in response to stimuli. We present a split ubiquitin-based method for detection of integral membrane protein-protein interactions...

  • Molecular interactions between mitochondrial membrane proteins and the C-terminal domain of PB1-F2: an in silico approach. Mohd Danishuddin; Shahper Khan; Asad Khan // Journal of Molecular Modeling;Mar2010, Vol. 16 Issue 3, p535 

    Abstract  PB1-F2 is a recently described influenza A viral protein that induces apoptosis by binding with two mitochondrial membrane proteins, i.e. VDAC1 (outer membrane) and ANT3 (inner membrane). Knowledge of this binding mechanism could provide insights that would aid in the...

  • Protein-Protein Interactions in Membranes. Armstrong, Clare L.; Sandqvist, Erik; Rheinstädter, Maikel C. // Protein & Peptide Letters;Apr2011, Vol. 18 Issue 4, p344 

    No abstract available.

  • Membrane Proteins, a Biophysical Perspective. Carrer, Dolores C. // Current Protein & Peptide Science;Dec2011, Vol. 12 Issue 8, p684 

    No abstract available.

  • The BRCA1 Breast Cancer Suppressor: Regulation of Transport, Dynamics, and Function at Multiple Subcellular Locations. Henderson, Beric R. // Scientifica;2012, p1 

    Inherited mutations in the BRCA1 gene predispose to a higher risk of breast/ovarian cancer. The BRCA1 tumor suppressor is a 1863 amino acid protein with multiple protein interaction domains that facilitate its roles in regulating DNA repair and maintenance, cell cycle progression, transcription,...

  • Systems biology: A plant membrane-linked interactome.  // Nature Methods;Jul2014, Vol. 11 Issue 7, p714 

    The article reports on the move of A. M. Jones and colleagues in using the established year split-ubiquitin assay to assess the interactions of more than 3,000 membrane proteins from Arabidopsis, a small flowering plant.

  • Characterization of the motion of membrane proteins using high-speed atomic force microscopy. Casuso, Ignacio; Khao, Jonathan; Chami, Mohamed; Paul-Gilloteaux, Perrine; Husain, Mohamed; Duneau, Jean-Pierre; Stahlberg, Henning; Sturgis, James N.; Scheuring, Simon // Nature Nanotechnology;Aug2012, Vol. 7 Issue 8, p525 

    For cells to function properly, membrane proteins must be able to diffuse within biological membranes. The functions of these membrane proteins depend on their position and also on protein-protein and protein-lipid interactions. However, so far, it has not been possible to study simultaneously...

  • The Free Energy Landscape of Dimerization of a Membrane Protein, NanC. Dunton, Thomas A.; Goose, Joseph E.; Gavaghan, David J.; Sansom, Mark S. P.; Osborne, James M. // PLoS Computational Biology;Jan2014, Vol. 10 Issue 1, p1 

    Membrane proteins are frequently present in crowded environments, which favour lateral association and, on occasions, two-dimensional crystallization. To better understand the non-specific lateral association of a membrane protein we have characterized the free energy landscape for the...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics