the cysteine-rich protein thimet oligopeptidase as a model of the structural requirements for s-glutathiolation and oxidative oligomerizationcysteine-rich蛋白甲拌磷oligopeptidase s-glutathiolation模型的结构要求和氧化寡聚化.pdf

The Cysteine-Rich Protein Thimet Oligopeptidase as a Model of the Structural Requirements for S- glutathiolation and Oxidative Oligomerization 1 ´ 2,4 1 2 3 Alberto Malvezzi , Patrıcia M. Higa , Antonia T.-do Amaral , Gustavo M. Silva , Fabio C. Gozzo , 4 4 1 5 2 Emer S. Ferro , Leandro M. Castro , Leandro de Rezende , Gisele Monteiro , Marilene Demasi * ´ ˜ ˜ ˜ ´ 1 Instituto de Quımica, Universidade de Sao Paulo, Sao Paulo-SP, Brazil, 2 Instituto Butantan, Sao Paulo-SP, Brazil, 3 Instituto de Quımica, Universidade Estadual de ˆ ´ ˜ ˜ ˆ ˆ Campinas, Campinas-SP, Brazil, 4 Instituto de Ciencias Biomedicas, Universidade de Sao Paulo, Sao Paulo-SP, Brazil, 5 Faculdade de Ciencias Farmaceuticas, Universidade ˜ ˜ de Sao Paulo, Sao Paulo-SP, Brazil Abstract Thimet oligopeptidase (EP24.15) is a cysteine-rich metallopeptidase containing fifteen Cys residues and no intra-protein disulfide bonds. Previous work on this enzyme revealed that the oxidative oligomerization of EP24.15 is triggered by S- glutathiolation at physiological GSSG levels (10–50 mM) via a mechanism based on thiol-disulfide exchange. In the present work, our aim was to identify EP24.15 Cys residues that are prone to S-glutathiolation and to determine which structural features in the cysteinyl