title: Design and characterization of a cleavage-resistant Annexin A1 mutant to control inflammation in the microvasculature.
abstract: Human polymorphonuclear leukocytes adhesion to endothelial cells during the early stage of inflammation leads to cell surface externalization of Annexin A1 (AnxA1), an effector of endogenous anti-inflammation. The antiadhesive properties of AnxA1 become operative to finely tune polymorphonuclear leukocytes transmigration to the site of inflammation. Membrane bound proteinase 3 (PR3) plays a key role in this microenvironment by cleaving the N terminus bioactive domain of AnxA1. In the present study, we generated a PR3-resistant human recombinant AnxA1-named superAnxA1 (SAnxA1)-and tested its in vitro and in vivo properties in comparison to the parental protein. SAnxA1 bound and activated formyl peptide receptor 2 in a similar way as the parental protein, while showing a resistance to cleavage by recombinant PR3. SAnxA1 retained anti-inflammatory activities in the murine inflamed microcirculation (leukocyte adhesion being the readout) and in skin trafficking model. When longer-lasting models of inflammation were applied, SAnxA1 displayed stronger anti-inflammatory effect over time compared with the parental protein. Together these results indicate that AnxA1 cleavage is an important process during neutrophilic inflammation and that controlling the balance between AnxA1/PR3 activities might represent a promising avenue for the discovery of novel therapeutic approaches.
authors: William Harvey Research Institute, Barts and The London School of Medicine, Queen Mary University of London, London, UK.
authors: Magali
authors: M
authors: Pederzoli-Ribeil
authors: Francesco
authors: F
authors: Maione
authors: Dianne
authors: D
authors: Cooper
authors: Adam
authors: A
authors: Al-Kashi
authors: Jesmond
authors: J
authors: Dalli
authors: Mauro
authors: M
authors: Perretti
authors: Fulvio
authors: F
authors: D'Acquisto
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017305:Annexin A1 / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000893:Anti-Inflammatory Agents / Q000378:metabolism; D002448:Cell Adhesion; D002450:Cell Communication; D002465:Cell Movement; D042783:Endothelial Cells / Q000378:metabolism; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D007249:Inflammation / Q000473:pathology* / Q000517:prevention & control*; D008297:Male; D051379:Mice; D055806:Microvessels / Q000378:metabolism / Q000473:pathology*; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism*; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism; D011485:Protein Binding; D015202:Protein Engineering; D044042:Receptors, Formyl Peptide / Q000378:metabolism; D044067:Receptors, Lipoxin / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D017305:Annexin A1; D000893:Anti-Inflammatory Agents; C105101:FPR2 protein, human; D050505:Mutant Proteins; D044042:Receptors, Formyl Peptide; D044067:Receptors, Lipoxin; D011994:Recombinant Proteins