title: The interaction of enolase-1 with caveolae-associated proteins regulates its subcellular localization.
abstract: Cell-surface-associated proteolysis plays a crucial role in embryonic development, monocyte/macrophage recruitment and tumour cell invasion. The glycolytic enzyme ENO-1 (enolase-1) is translocated from the cytoplasm to the cell surface, where it binds PLG (plasminogen) to enhance pericellular plasmin production and cell motility. In the present study, ENO-1 was found to localize to a specialized subset of lipid rafts called caveolae as demonstrated by fluorescence confocal microscopy and sucrose gradient ultracentrifugation. Co-immunoprecipitation studies revealed that ENO-1 interacts with Cav-1 (caveolin-1), but not with Cav-2, via the CSD (Cav-scaffolding domain). Moreover, an evolutionarily conserved CBM (Cav-binding motif) F296DQDDWGAW304 was identified within ENO-1. The point mutation W301A within the ENO-1 CBM was, however, not sufficient to disrupt ENO-1-Cav-1 interaction, whereas the mutations F296A and W304A markedly affected ENO-1 protein expression. Furthermore, ENO-1 was found associated with Annx2 (annexin 2), representing another caveolar protein, and this interaction was dependent on Cav-1 expression. Knockdown of Cav-1 and Annx2 markedly decreased cell surface expression of ENO-1. ENO-1 overexpression increased cell migration and invasion in a Cav-1-dependent manner. Thus the differential association of ENO-1 with caveolar proteins regulates ENO-1 subcellular localization and, consequently, ENO-1-dependent cell migration and invasion.
authors.affiliation: *Department of Biochemistry, Faculty of Medicine, University of Giessen Lung Center, Friedrichstrasse 24, 35392 Giessen, Germany.
authors.forename: Dariusz
authors.initials: D
authors.lastname: Zakrzewicz
authors.affiliation: *Department of Biochemistry, Faculty of Medicine, University of Giessen Lung Center, Friedrichstrasse 24, 35392 Giessen, Germany.
authors.forename: Miroslava
authors.initials: M
authors.lastname: Didiasova
authors.affiliation: †Laboratory of Experimental Surgery, Department of General and Thoracic Surgery, Justus Liebig-University Giessen, Feulgenstrasse 10-12, 35385 Giessen, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Zakrzewicz
authors.affiliation: ‡Department of Internal Medicine/Infectious Diseases and Pulmonary Medicine, Charité-University Medicine Berlin, Chariteplatz 1, 10117 Berlin, Germany.
authors.forename: Andreas C
authors.initials: AC
authors.lastname: Hocke
authors.affiliation: §Departments of Anaesthesiology and Operative Intensive Care, University Hospital Giessen and Marburg GmbH, Klinikstrasse 33, Giessen 35392, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Uhle
authors.forename: Philipp
authors.initials: P
authors.lastname: Markart
authors.forename: Klaus T
authors.initials: KT
authors.lastname: Preissner
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Wygrecka
mesh_terms: D000818:Animals; D017306:Annexin A2 / Q000378:metabolism*; D014408:Biomarkers, Tumor / Q000378:metabolism*; D021941:Caveolae / Q000378:metabolism*; D051242:Caveolin 1 / Q000378:metabolism*; D002465:Cell Movement; D004268:DNA-Binding Proteins / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D010751:Phosphopyruvate Hydratase / Q000378:metabolism*; D010958:Plasminogen / Q000378:metabolism; D021381:Protein Transport; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins / Q000378:metabolism*
chemical_list: C472822:ANXA2 protein, human; D017306:Annexin A2; D014408:Biomarkers, Tumor; D051242:Caveolin 1; D004268:DNA-Binding Proteins; D025521:Tumor Suppressor Proteins; D010958:Plasminogen; C095157:ENO1 protein, human; D010751:Phosphopyruvate Hydratase