PIMT is a novel and potent suppressor of endothelial activation

Proinflammatory agonists provoke the expression of cell surface adhesion molecules on endothelium in order to facilitate leukocyte infiltration into tissues. Rigorous control over this process is important to prevent unwanted inflammation and organ damage. Protein L-isoaspartyl O-methyltransferase (PIMT) converts isoaspartyl residues to conventional methylated forms in cells undergoing stress-induced protein damage. The purpose of this study was to determine the role of PIMT in vascular homeostasis. PIMT is abundantly expressed in mouse lung endothelium and PIMT deficiency in mice exacerbated pulmonary inflammation and vascular leakage to LPS(lipopolysaccharide). Furthermore, we found that PIMT inhibited LPS-induced toll-like receptor signaling through its interaction with TNF receptor-associated factor 6 (TRAF6) and its ability to methylate asparagine residues in the coiled-coil domain. This interaction was found to inhibit TRAF6 oligomerization and autoubiquitination, which prevented NF-κB transactivation and subsequent expression of endothelial adhesion molecules. Separately, PIMT also suppressed ICAM-1 expression by inhibiting its N-glycosylation, causing effects on protein stability that ultimately translated into reduced EC(endothelial cell)-leukocyte interactions. Our study has identified PIMT as a novel and potent suppressor of endothelial activation. Taken together, these findings suggest that therapeutic targeting of PIMT may be effective in limiting organ injury in inflammatory vascular diseases.

[1]  D. Gilroy,et al.  Aging immunity may exacerbate COVID-19 , 2020, Science.

[2]  Sook-In Jung,et al.  Age-Related Morbidity and Mortality among Patients with COVID-19 , 2020, Infection & chemotherapy.

[3]  Wenjie Zhu,et al.  Deamidation Shunts RelA from Mediating Inflammation to Aerobic Glycolysis. , 2020, Cell metabolism.

[4]  R. Summer,et al.  Nur77 limits endothelial barrier disruption to LPS in the mouse lung. , 2019, American journal of physiology. Lung cellular and molecular physiology.

[5]  M. Mahawar,et al.  PIMT-Mediated Protein Repair: Mechanism and Implications , 2019, Biochemistry (Moscow).

[6]  Wenlin Huang,et al.  Cyclophilin J limits inflammation through the blockage of ubiquitin chain sensing , 2018, Nature Communications.

[7]  G. Hu,et al.  YAP Controls Endothelial Activation and Vascular Inflammation Through TRAF6 , 2018, Circulation research.

[8]  D. Neculai,et al.  Oligomerization-primed coiled-coil domain interaction with Ubc13 confers processivity to TRAF6 ubiquitin ligase activity , 2017, Nature Communications.

[9]  M. Kumawat,et al.  Protein-L-Isoaspartyl Methyltransferase (PIMT) Is Required for Survival of Salmonella Typhimurium at 42°C and Contributes to the Virulence in Poultry , 2017, Front. Microbiol..

[10]  M. Griffiths,et al.  The pulmonary endothelium in acute respiratory distress syndrome: insights and therapeutic opportunities , 2016, Thorax.

[11]  Amira Ouanouki,et al.  The enzyme l-isoaspartyl (d-aspartyl) methyltransferase is required for VEGF-dependent endothelial cell migration and tubulogenesis , 2016, Molecular and Cellular Biochemistry.

[12]  R. Schneider,et al.  Methylation of histone H4 at aspartate 24 by Protein L-isoaspartate O-methyltransferase (PCMT1) links histone modifications with protein homeostasis , 2014, Scientific Reports.

[13]  Kou-Juey Wu,et al.  Twist1 induces endothelial differentiation of tumour cells through the Jagged1-KLF4 axis , 2014, Nature Communications.

[14]  Jianxin Sun,et al.  Inhibition of Cardiomyocyte Hypertrophy by Protein Arginine Methyltransferase 5* , 2014, The Journal of Biological Chemistry.

[15]  L. Dang,et al.  Aberrant IKKα and IKKβ cooperatively activate NF-κB and induce EGFR/AP1 signaling to promote survival and migration of head and neck cancer , 2013, Oncogene.

[16]  G. Yan,et al.  Protein-L-isoaspartate (D-aspartate) O-methyltransferase protects cardiomyocytes against hypoxia induced apoptosis through inhibiting proapoptotic kinase Mst1. , 2013, International journal of cardiology.

[17]  D. Scott,et al.  Endothelial heterogeneity and adhesion molecules N-glycosylation: implications in leukocyte trafficking in inflammation. , 2013, Glycobiology.

[18]  S. Clarke,et al.  Integrated proteomic analysis of major isoaspartyl-containing proteins in the urine of wild type and protein L-isoaspartate O-methyltransferase-deficient mice. , 2013, Analytical chemistry.

[19]  Ju-Seog Lee,et al.  Protein L-isoaspartyl methyltransferase regulates p53 activity , 2012, Nature Communications.

[20]  R. Desrosiers,et al.  Damaged proteins bearing L-isoaspartyl residues and aging: a dynamic equilibrium between generation of isomerized forms and repair by PIMT. , 2011, Current aging science.

[21]  Shaogang Sun,et al.  Negative Feedback Regulation of NF-κB Action by CITED2 in the Nucleus , 2011, The Journal of Immunology.

[22]  Alexander Hoffmann,et al.  The regulatory logic of the NF-kappaB signaling system. , 2010, Cold Spring Harbor perspectives in biology.

[23]  T. Lawrence.  The nuclear factor NF-kappaB pathway in inflammation. , 2009, Cold Spring Harbor perspectives in biology.

[24]  Zhuohua Zhang,et al.  Detection of protein ubiquitination. , 2009, Journal of visualized experiments : JoVE.

[25]  Greg L. Hura,et al.  E2 interaction and dimerization in the crystal structure of TRAF6 , 2009, Nature Structural &Molecular Biology.

[26]  C. Lawson,et al.  ICAM-1 signaling in endothelial cells , 2009, Pharmacological reports : PR.

[27]  D. Job,et al.  Protein Repair l-Isoaspartyl Methyltransferase1 Is Involved in Both Seed Longevity and Germination Vigor in Arabidopsis[W] , 2008, The Plant Cell Online.

[28]  J. Catravas,et al.  Endothelial pathomechanisms in acute lung injury , 2008, Vascular Pharmacology.

[29]  M. Wurfel,et al.  A TRIFfic Perspective on Acute Lung Injury , 2008, Cell.

[30]  Kai Zhang,et al.  Identification and validation of eukaryotic aspartate and glutamate methylation in proteins. , 2008, Journal of proteome research.

[31]  Jordan S. Pober,et al.  Evolving functions of endothelial cells in inflammation , 2007, Nature Reviews Immunology.

[32]  G. Pei,et al.  Association of β-arrestin and TRAF6 negatively regulates Toll-like receptor–interleukin 1 receptor signaling , 2006, Nature Immunology.

[33]  A. Pries,et al.  Normal endothelium. , 2006, Handbook of experimental pharmacology.

[34]  A. Karsan,et al.  Lipopolysaccharide signaling in endothelial cells , 2006, Laboratory Investigation.

[35]  Zhijian J. Chen.  Ubiquitin signalling in the NF-κB pathway , 2005, Nature Cell Biology.

[36]  Matthew T Wheeler,et al.  The ubiquitin-modifying enzyme A20 is required for termination of Toll-like receptor responses , 2004, Nature Immunology.

[37]  C. Roussos,et al.  Pulmonary endothelium in acute lung injury: from basic science to the critically ill , 2004, Intensive Care Medicine.

[38]  Shizuo Akira,et al.  Toll-like receptor signalling , 2004, Nature Reviews Immunology.

[39]  N R Webster,et al.  Physiology of the endothelium. , 2004, British journal of anaesthesia.

[40]  Jia-Huai Wang,et al.  Structural basis for dimerization of ICAM-1 on the cell surface. , 2004, Molecular cell.

[41]  S. Verma,et al.  New Markers of Inflammation and Endothelial Cell Activation: Part I , 2003, Circulation.

[42]  D. Aswad,et al.  Deamidation and isoaspartate formation in proteins: unwanted alterations or surreptitious signals? , 2003, Cellular and Molecular Life Sciences CMLS.

[43]  Junichi Takagi,et al.  Structures of the αL I Domain and Its Complex with ICAM-1 Reveal a Shape-Shifting Pathway for Integrin Regulation , 2003, Cell.

[44]  A. Dunne,et al.  The Interleukin-1 Receptor/Toll-Like Receptor Superfamily: Signal Transduction During Inflammation and Host Defense , 2000, Science's STKE.

[45]  Ellen M. Langer,et al.  Bcl-xL Deamidation Is a Critical Switch in the Regulation of the Response to DNA Damage , 2002, Cell.

[46]  Hao Wu,et al.  Distinct molecular mechanism for initiating TRAF6 signalling , 2002, Nature.

[47]  Hsien-Yeh Hsu,et al.  Lipopolysaccharide-mediated Reactive Oxygen Species and Signal Transduction in the Regulation of Interleukin-1 Gene Expression* , 2002, The Journal of Biological Chemistry.

[48]  Young Chul Park,et al.  All TRAFs are not created equal: common and distinct molecular mechanisms of TRAF-mediated signal transduction. , 2002, Journal of cell science.

[49]  F. Jackson,et al.  Extension of the Drosophila lifespan by overexpression of a protein repair methyltransferase , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[50]  Zhijian J. Chen,et al.  TAK1 is a ubiquitin-dependent kinase of MKK and IKK , 2001, Nature.

[51]  B. Perfetto,et al.  UVA irradiation induces L-isoaspartyl formation in melanoma cell proteins. , 2001, Free radical biology & medicine.

[52]  Zhijian J. Chen,et al.  Activation of the IκB Kinase Complex by TRAF6 Requires a Dimeric Ubiquitin-Conjugating Enzyme Complex and a Unique Polyubiquitin Chain , 2000, Cell.

[53]  P. Bates,et al.  The second domain of intercellular adhesion molecule-1 (ICAM-1) maintains the structural integrity of the leucocyte function-associated antigen-1 (LFA-1) ligand-binding site in the first domain. , 2000, The Biochemical journal.

[54]  T. Muta,et al.  TAK1 mediates an activation signal from toll‐like receptor(s) to nuclear factor‐κB in lipopolysaccharide‐stimulated macrophages , 2000, FEBS letters.

[55]  M. Yacoub,et al.  Ligation of ICAM-1 on endothelial cells leads to expression of VCAM-1 via a nuclear factor-kappaB-independent mechanism. , 1999, Journal of immunology.

[56]  P. Baeuerle.  IκB–NF-κB Structures At the Interface of Inflammation Control , 1998, Cell.

[57]  L. Kasturi,et al.  The amino acid following an asn-X-Ser/Thr sequon is an important determinant of N-linked core glycosylation efficiency. , 1998, Biochemistry.

[58]  B. Hunt,et al.  Endothelial cell activation , 1998, BMJ.

[59]  T. Springer,et al.  A dimeric crystal structure for the N-terminal two domains of intercellular adhesion molecule-1. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[60]  S. Young,et al.  Deficiency of a protein-repair enzyme results in the accumulation of altered proteins, retardation of growth, and fatal seizures in mice. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[61]  K. Beningo,et al.  A permanent human cell line (EA.hy926) preserves the characteristics of endothelin converting enzyme from primary human umbilical vein endothelial cells. , 1995, Life sciences.

[62]  R. Kagan,et al.  Widespread occurrence of three sequence motifs in diverse S-adenosylmethionine-dependent methyltransferases suggests a common structure for these enzymes. , 1994, Archives of biochemistry and biophysics.

[63]  T. Springer.  Traffic signals for lymphocyte recirculation and leukocyte emigration: The multistep paradigm , 1994, Cell.

[64]  M. Diamond,et al.  The I domain is a major recognition site on the leukocyte integrin Mac- 1 (CD11b/CD18) for four distinct adhesion ligands , 1993, The Journal of cell biology.

[65]  Michael Loran Dustin,et al.  Primary structure of ICAM-1 demonstrates interaction between members of the immunoglobulin and integrin supergene families , 1988, Cell.

[66]  S. Clarke,et al.  Deamidation, isomerization, and racemization at asparaginyl and aspartyl residues in peptides. Succinimide-linked reactions that contribute to protein degradation. , 1987, The Journal of biological chemistry.