title: Neutrophil elastase cleavage of the gC1q domain impairs the EMILIN1-α4β1 integrin interaction, cell adhesion and anti-proliferative activity.
abstract: The extracellular matrix glycoprotein EMILIN1 exerts a wide range of functions mainly associated with its gC1q domain. Besides providing functional significance for adhesion and migration, the direct interaction between α4β1 integrin and EMILIN1-gC1q regulates cell proliferation, transducing net anti-proliferative effects. We have previously demonstrated that EMILIN1 degradation by neutrophil elastase (NE) is a specific mechanism leading to the loss of functions disabling its regulatory properties. In this study we further analysed the proteolytic activity of NE, MMP-3, MMP-9, and MT1-MMP on EMILIN1 and found that MMP-3 and MT1-MMP partially cleaved EMILIN1 but without affecting the functional properties associated with the gC1q domain, whereas NE was able to fully impair the interaction of gC1q with the α4β1 integrin by cleaving this domain outside of the E933 integrin binding site. By a site direct mutagenesis approach we mapped the bond between S913 and R914 residues and selected the NE-resistant R914W mutant still able to interact with the α4β1 integrin after NE treatment. Functional studies showed that NE impaired the EMILIN1-α4β1 integrin interaction by cleaving the gC1q domain in a region crucial for its proper structural conformation, paving the way to better understand NE effects on EMILIN1-cell interaction in pathological context.
authors.affiliation: Experimental Oncology 2, Department of Translational Research, CRO-IRCCS, National Cancer Institute, Aviano 33081, Italy.
authors.forename: Orlando
authors.initials: O
authors.lastname: Maiorani
authors.affiliation: Experimental Oncology 2, Department of Translational Research, CRO-IRCCS, National Cancer Institute, Aviano 33081, Italy.
authors.forename: Eliana
authors.initials: E
authors.lastname: Pivetta
authors.affiliation: Experimental Oncology 2, Department of Translational Research, CRO-IRCCS, National Cancer Institute, Aviano 33081, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Capuano
authors.affiliation: Experimental Oncology 2, Department of Translational Research, CRO-IRCCS, National Cancer Institute, Aviano 33081, Italy.
authors.forename: Teresa Maria Elisa
authors.initials: TM
authors.lastname: Modica
authors.affiliation: Experimental Oncology 2, Department of Translational Research, CRO-IRCCS, National Cancer Institute, Aviano 33081, Italy.
authors.forename: Bruna
authors.initials: B
authors.lastname: Wassermann
authors.affiliation: Experimental Oncology 2, Department of Translational Research, CRO-IRCCS, National Cancer Institute, Aviano 33081, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Bucciotti
authors.affiliation: Experimental Oncology 2, Department of Translational Research, CRO-IRCCS, National Cancer Institute, Aviano 33081, Italy.
authors.forename: Alfonso
authors.initials: A
authors.lastname: Colombatti
authors.affiliation: Experimental Oncology 2, Department of Translational Research, CRO-IRCCS, National Cancer Institute, Aviano 33081, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Doliana
authors.affiliation: Experimental Oncology 2, Department of Translational Research, CRO-IRCCS, National Cancer Institute, Aviano 33081, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Spessotto
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D020134:Catalytic Domain; D002448:Cell Adhesion / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D039041:Integrin alpha4beta1 / Q000737:chemistry / Q000378:metabolism*; D019272:Leukocyte Elastase / Q000378:metabolism*; D053511:Matrix Metalloproteinase 14 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019278:Matrix Metalloproteinase 3 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D059748:Proteolysis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: C087299:C1QBP protein, human; D002352:Carrier Proteins; D039041:Integrin alpha4beta1; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; D011994:Recombinant Proteins; C030340:elastin microfibril interface located protein; D019272:Leukocyte Elastase; D019278:Matrix Metalloproteinase 3; D020780:Matrix Metalloproteinase 9; D053511:Matrix Metalloproteinase 14