title: Role of protein-tyrosine phosphatase SHP2 in focal adhesion kinase down-regulation during neutrophil cathepsin G-induced cardiomyocytes anoikis.
abstract: Inflammatory cells and their proteases contribute to tissue reparation at site of inflammation. Although beneficial at early stages, excessive inflammatory reaction leads to cell death and tissue damage. Cathepsin G (Cat.G), a neutrophil-derived serine protease, has been shown to induce neonatal rat cardiomyocyte detachment and apoptosis by anoikis through caspase-3 dependent pathway. However the early mechanisms that trigger Cat.G-induced caspase-3 activation are not known. This study identifies focal adhesion kinase (FAK) tyrosine dephosphorylation as an early mechanism that regulates Cat.G-induced anoikis in cardiomyocytes. Both FAK tyrosine phosphorylation at Tyr-397 and kinase activity decrease rapidly upon Cat.G treatment and was associated with a decrease of FAK association with adapter and cytoskeletal proteins, p130(Cas) and paxillin, respectively. FAK-decreased tyrosine phosphorylation is required for Cat.G-induced myocyte anoikis as concurrent expression of phosphorylation-deficient FAK mutated at Tyr-397 or pretreatment with a protein-tyrosine phosphatase (PTP) inhibitor, pervanadate, blocks Cat.G-induced FAK tyrosine dephosphorylation, caspase-3 activation and DNA fragmentation. Analysis of PTPs activation shows that Cat.G treatment induces an increase of SHP2 and PTEN phosphorylation; however, only SHP2 forms a complex with FAK in response to Cat.G. Expression of dominant negative SHP2 mutant markedly attenuates FAK tyrosine dephosphorylation induced by Cat.G and protects myocytes to undergo apoptosis. In contrast, increased SHP2 expression exacerbates Cat.G-induced FAK tyrosine dephosphorylation and myocyte apoptosis. Taken together, these results show that Cat.G induces SHP2 activation that leads to FAK tyrosine dephosphorylation and promotes cardiomyocyte anoikis.
authors.affiliation: Cardiovascular Research Center, Department of Anatomy and Cell Biology, Temple University, Philadelphia, Pennsylvania 19140, USA.
authors.forename: Khadija
authors.initials: K
authors.lastname: Rafiq
authors.forename: Mikhail A
authors.initials: MA
authors.lastname: Kolpakov
authors.forename: Malika
authors.initials: M
authors.lastname: Abdelfettah
authors.forename: Daniel N
authors.initials: DN
authors.lastname: Streblow
authors.forename: Aviv
authors.initials: A
authors.lastname: Hassid
authors.forename: Louis J
authors.initials: LJ
authors.lastname: Dell'Italia
authors.forename: Abdelkarim
authors.initials: A
authors.lastname: Sabri
mesh_terms: D000256:Adenoviridae / Q000378:metabolism; D000818:Animals; D023102:Anoikis; D017209:Apoptosis; D056649:Cathepsin G; D002403:Cathepsins / Q000378:metabolism*; D015536:Down-Regulation; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry / Q000502:physiology*; D032383:Myocytes, Cardiac / Q000378:metabolism*; D009504:Neutrophils / Q000378:metabolism*; D010766:Phosphorylation; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012697:Serine Endopeptidases / Q000378:metabolism*; D013347:Subcellular Fractions
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D051416:Focal Adhesion Protein-Tyrosine Kinases; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D017027:Protein Tyrosine Phosphatases; C516727:Ptpn11 protein, rat; D002403:Cathepsins; D012697:Serine Endopeptidases; D056649:Cathepsin G; C534097:Ctsg protein, rat