Abstract
Several experimental approaches have demonstrated that transglutaminase 2 (TG2) increased activity is involved in monocyte activation and inflammatory response. Preliminary results also demonstrate a TG-mediated post-translational modification of phospholipase A2 (PLA2), which catalyzes the release of arachidonic acid from its lipid storage sites. The control of PLA2-mediated production of eicosanoids has been found to be of great benefit for inflammatory disease treatment. However, the identification of the mechanisms of PLA2 activation is a very complex issue, because of the presence of multiple PLA2 forms. The aim of this study was to characterize the interactions between TG2 and sPLA2 in LPS-stimulated THP-1 cells, which were treated with TPA to induce early differentiated macrophage-type model. We demonstrated that increases in TG2 enzyme activity and protein expression may be considered an early event in monocyte/macrophage activation by LPS. Under these conditions, TG2 protein was co-immunoprecipitated with PLA2 by monoclonal antibody directed against the secretory form of the enzyme (sPLA2-V). Concomitantly, the PLA2 enzyme activity increased in TPA-treated cells exposed to LPS; these high levels of enzyme activity were significant reduced by R283, a site-specific inhibitor of TG2. Moreover, confocal laser scanning microscopy analysis of double-immunostained cytochemical specimens confirmed a co-localization of BAPA-labeled proteins and sPLA2-V in LPS-treated cells. These findings give evidence of a complex TG2/sPLA2-V, suggesting the possibility that sPLA2-V is a substrate for TG2. These results demonstrated that TG2 increases produced a sustained activation of PLA2 activity, suggesting a functional interaction between these enzymes in the regulation of inflammatory response.
Similar content being viewed by others
References
Aeschlimann D, Paulsson M (1994) Transglutaminases: protein cross-linking enzymes in tissues and body fluids. Thromb Haemost 71:402–415
Baek SH, Kwon TK, Lim JH et al (2000) Secretory phospholipase A2-potentiated inducible nitric oxide synthase expression by macrophages requires NF-kappa B activation. J Immunol 164:6359–6365
Cervellati C, Montin K, Squerzanti M et al (2012) Effects of the regulatory ligands calcium and GTP on the thermal stability of tissue transglutaminase. Amino Acids 42:2233–2242. doi:10.1007/s00726-011-0963-6
Chakraborti S (2003) Phospholipase A(2) isoforms: a perspective. Cell Signal 15:637–665
Cordella-Miele E, Miele L, Mukherjee AB (1990) A novel transglutaminase-mediated post-translational modification of phospholipase A2 dramatically increases its catalytic activity. J Biol Chem 265:17180–17188
Cordella-Miele E, Miele L, Beninati S, Mukherjee AB (1993) Transglutaminase-catalyzed incorporation of polyamines into phospholipase A2. J Biochem 113:164–173
Currò M, Ferlazzo N, Condello S et al (2010) Transglutaminase 2 silencing reduced the beta-amyloid-effects on the activation of human THP-1 cells. Amino Acids 39:1427–1433. doi:10.1007/s00726-010-0605-4
Fink ML, Folk JE (1981) Gamma-glutamylamine cyclotransferase. An enzyme involved in the catabolism of epsilon-(gamma-glutamyl) lysine and other gamma-glutamylamines. Mol Cell Biochem 38:59–67
Garabuczi É, Kiss B, Felszeghy S et al (2013) Retinoids produced by macrophages engulfing apoptotic cells contribute to the appearance of transglutaminase 2 in apoptotic thymocytes. Amino Acids 44:235–244. doi:10.1007/s00726-011-1119-4
Griffin M, Casadio R, Bergamini CM (2002) Transglutaminases: nature’s biological glues. Biochem J 368:377–396. doi:10.1042/BJ20021234
Ientile R, Caccamo D, Griffin M (2007) Tissue transglutaminase and the stress response. Amino Acids 33:385–394. doi:10.1007/s00726-007-0517-0
Jeitner TM, Pinto JT, Krasnikov BF et al (2009) Transglutaminases and neurodegeneration. J Neurochem 109:160–166. doi:10.1111/j.1471-4159.2009.05843.x
Kim SY (2006) Transglutaminase 2 in inflammation. Front Biosci J Virtual Libr 11:3026–3035
Kim SY, Jeitner TM, Steinert PM (2002) Transglutaminases in disease. Neurochem Int 40:85–103
Király R, Demény M, Fésüs L (2011) Protein transamidation by transglutaminase 2 in cells: a disputed Ca2+-dependent action of a multifunctional protein. FEBS J 278:4717–4739. doi:10.1111/j.1742-4658.2011.08345.x
Krig SR, Chandraratna RAS, Chang MMJ et al (2002) Gene-specific TCDD suppression of RARalpha- and RXR-mediated induction of tissue transglutaminase. Toxicol Sci Off J Soc Toxicol 68:102–108
Kumar S, Mehta K (2012) Tissue transglutaminase constitutively activates HIF-1α promoter and nuclear factor-κB via a non-canonical pathway. PLoS ONE 7:e49321. doi:10.1371/journal.pone.0049321
Mehta K, Kumar A, Kim HI (2010) Transglutaminase 2: a multi-tasking protein in the complex circuitry of inflammation and cancer. Biochem Pharmacol 80:1921–1929. doi:10.1016/j.bcp.2010.06.029
Mirza A, Liu SL, Frizell E et al (1997) A role for tissue transglutaminase in hepatic injury and fibrogenesis, and its regulation by NF-kappaB. Am J Physiol 272:G281–G288
Moreno JJ (2006) Effects of antiflammins on transglutaminase and phospholipase A2 activation by transglutaminase. Int Immunopharmacol 6:300–303. doi:10.1016/j.intimp.2005.08.001
Murtaugh MP, Mehta K, Johnson J et al (1983) Induction of tissue transglutaminase in mouse peritoneal macrophages. J Biol Chem 258:11074–11081
Novogrodsky A, Quittner S, Rubin AL, Stenzel KH (1978) Transglutaminase activity in human lymphocytes: early activation by phytomitogens. Proc Natl Acad Sci U S A 75:1157–1161
Park D, Choi SS, Ha K-S (2010) Transglutaminase 2: a multi-functional protein in multiple subcellular compartments. Amino Acids 39:619–631. doi:10.1007/s00726-010-0500-z
Qin Z (2012) The use of THP-1 cells as a model for mimicking the function and regulation of monocytes and macrophages in the vasculature. Atherosclerosis 221:2–11. doi:10.1016/j.atherosclerosis.2011.09.003
Quan G, Choi J-Y, Lee D-S, Lee S-C (2005) TGF-beta1 up-regulates transglutaminase two and fibronectin in dermal fibroblasts: a possible mechanism for the stabilization of tissue inflammation. Arch Dermatol Res 297:84–90. doi:10.1007/s00403-005-0582-8
Schroff G, Neumann C, Sorg C (1981) Transglutaminase as a marker for subsets of murine macrophages. Eur J Immunol 11:637–642. doi:10.1002/eji.1830110809
Seiving B, Ohlsson K, Linder C, Stenberg P (1991) Transglutaminase differentiation during maturation of human blood monocytes to macrophages. Eur J Haematol 46:263–271
Szondy Z, Korponay-Szabó I, Király R, Fésüs L (2011) Transglutaminase 2 dysfunctions in the development of autoimmune disorders: celiac disease and TG2-/- mouse. Adv Enzymol Relat Areas Mol Biol 78:295–345
Turner PM, Lorand L (1989) Complexation of fibronectin with tissue transglutaminase. Biochemistry 28:628–635
Verhaar R, Jongenelen CAM, Gerard M et al (2011) Blockade of enzyme activity inhibits tissue transglutaminase-mediated transamidation of α-synuclein in a cellular model of Parkinson’s disease. Neurochem Int 58:785–793. doi:10.1016/j.neuint.2011.03.004
Wang Z, Griffin M (2012) TG2, a novel extracellular protein with multiple functions. Amino Acids 42:939–949. doi:10.1007/s00726-011-1008-x
Zhang J, Lesort M, Guttmann RP, Johnson GV (1998) Modulation of the in situ activity of tissue transglutaminase by calcium and GTP. J Biol Chem 273:2288–2295
Acknowledgments
We thank Prof. M. Griffin (Aston University, Birmingham, UK) for the gift of the TG2-specific inhibitor R283.
Conflict of interest
The authors disclose no potential conflicts of interest.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Currò, M., Ferlazzo, N., Risitano, R. et al. Transglutaminase 2 and phospholipase A2 interactions in the inflammatory response in human Thp-1 monocytes. Amino Acids 46, 759–766 (2014). https://doi.org/10.1007/s00726-013-1569-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00726-013-1569-y