title: Monoclonal antibodies against tyrosyl-tRNA synthetase and its isolated cytokine-like domain.
abstract: Tyrosyl-tRNA synthetase (TyrRS) is one of the key enzymes of protein biosynthesis. In addition to its basic role, this enzyme reveals some important non-canonical functions. Under apoptotic conditions, the full-length enzyme splits into two fragments having distinct cytokine activities, thereby linking protein synthesis to cytokine signaling pathways. The NH2-terminal catalytic fragment, known as miniTyrRS, binds strongly to the CXC-chemokine receptor CXCR1 and, like interleukin 8, functions as a chemoattractant for polymorphonuclear leukocytes. On the other hand, an extra COOH-terminal domain of human TyrRS has cytokine activities like those of a mature human endothelial monocyte-activating polypeptide II (EMAP II). Moreover, the etiology of specific diseases (cancer, neuronal pathologies, autoimmune disorders, and disrupted metabolic conditions) is connected to specific aminoacyl-tRNA synthetases. Here we report the generation and characterization of monoclonal antibodies specific to N- and C-terminal domains of TyrRS. Recombinant TyrRS and its N- and C-terminal domains were expressed as His-tag fusion proteins in bacteria. Affinity purified proteins have been used as antigens for immunization and hybridoma cell screening. Monoclonal antibodies specific to catalytic N-terminal module and C-terminal EMAP II-like domain of TyrRS may be useful as tools in various aspects of TyrRS function and cellular localization.
authors.affiliation: Department of Protein Engineering and Bioinformatics, Institute of Molecular Biology and Genetics, National Academy of Science of Ukraine, Kyiv, Ukraine. kondratyuk_yulya@ukr.net
authors.forename: Iuliia
authors.initials: I
authors.lastname: Kondratiuk
authors.forename: Antonina
authors.initials: A
authors.lastname: Khoruzenko
authors.forename: Olga
authors.initials: O
authors.lastname: Cherednyk
authors.forename: Valeriy
authors.initials: V
authors.lastname: Filonenko
authors.forename: Aleksander
authors.initials: A
authors.lastname: Kornelyuk
mesh_terms: D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D020134:Catalytic Domain / Q000276:immunology*; D002460:Cell Line; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000276:immunology*; D014448:Tyrosine-tRNA Ligase / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D014448:Tyrosine-tRNA Ligase