118921 20240319081008.0 doi 10.3390/ijms23158705 sideral 130035 ART-2022-130035 eng Soler Agesta, R. Universidad de Zaragoza Conjugation of the 9-kDa isoform of Granulysin with liposomes potentiates its cytotoxicity 2022 Access copy available to the general public Unrestricted Nine kDa granulysin (GRNLY) is a human cytolytic protein secreted by cytotoxic T lymphocytes (CTL) and NK cells of the immune system whose demonstrated physiological function is the elimination of bacteria and parasites. In previous studies by our group, the anti-tumor capacity of recombinant granulysin was demonstrated, both in vitro and in vivo. In the present work, we developed lipid nanoparticles whose surfaces can bind recombinant granulysin through the formation of a complex of coordination between the histidine tail of the protein and Ni2+ provided by a chelating lipid in the liposome composition and termed them LUV-GRNLY, for granulysin-bound large unilamellar vesicles. The objective of this formulation is to increase the granulysin concentration at the site of contact with the target cell and to increase the cytotoxicity of the administered dose. The results obtained in this work indicate that recombinant granulysin binds to the surface of the liposome with high efficiency and that its cytotoxicity is significantly increased when it is in association with liposomes. In addition, it has been demonstrated that the main mechanism of death induced by both granulysin and LUV-GRNLY is apoptosis. Jurkat-shBak cells are resistant to GRNLY and also to LUV-GRNLY, showing that LUV-GRNLY uses the mitochondrial apoptotic pathway to induce cell death. On the other hand, we show that LUV-GRNLY induces the expression of the pro-apoptotic members of the Bcl-2 family Bim and especially PUMA, although it also induced the expression of anti-apoptotic Bcl-xL. In conclusion, we demonstrate that binding of GRNLY to the surfaces of liposomes clearly augments its cytotoxic potential, with cell death executed mainly by the mitochondrial apoptotic pathway. info:eu-repo/grantAgreement/ES/DGA/B31-20R info:eu-repo/grantAgreement/ES/MICINN/PID2019-105128RB-I00 info:eu-repo/semantics/openAccess by http://creativecommons.org/licenses/by/3.0/es/ 5.6 2022 BIOCHEMISTRY & MOLECULAR BIOLOGY 66 / 285 = 0.232 2022 Q1 T1 CHEMISTRY, MULTIDISCIPLINARY 52 / 178 = 0.292 2022 Q2 T1 1.154 2022 Medicine (miscellaneous) 2022 Q1 Physical and Theoretical Chemistry 2022 Q1 Computer Science Applications 2022 Q1 Inorganic Chemistry 2022 Q1 Spectroscopy 2022 Q1 Organic Chemistry 2022 Q1 Molecular Biology 2022 Q2 Catalysis 2022 Q2 7.8 2022 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Guerrero-Ochoa, P. Universidad de Zaragoza (orcid)0000-0002-1657-4792 Marco-Brualla, J. Universidad de Zaragoza Ibáñez-Pérez, R. Marzo, I. Universidad de Zaragoza (orcid)0000-0002-2315-9079 Martínez Lostao, L. Universidad de Zaragoza (orcid)0000-0003-3043-147X Anel, A. Universidad de Zaragoza (orcid)0000-0002-5175-8394 1011 566 Universidad de Zaragoza Dpto. Microb.Ped.Radio.Sal.Pú. Área Inmunología 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 1002 050 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Biología Celular 23, 15 (2022), 8705 [13 pp.] Int. j. mol. sci. International Journal of Molecular Sciences 1661-6596 1981037 http://zaguan.unizar.es/record/118921/files/texto_completo.pdf Versión publicada 2649865 http://zaguan.unizar.es/record/118921/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:118921 articulos driver 2024-03-18-14:50:13 ARTICLE