Diagnostic test for ruthenium and platinum modification of histidine residues on metalloproteins using diethylpyrocarbonate (DEPC) |
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| Institution: | 1. “Atmospheric Pollution” Associate Unit, CSIC-University of Huelva, Centre for Research in Sustainable Chemistry-CIQSO, University of Huelva, Campus de El Carmen s/n, 21071, Huelva, Spain;2. Department of Civil and Environmental, Universidad de la Costa, Calle 58 #55-66, 080002, Barranquilla, Colombia;3. Department of Earth Sciences, University of Huelva, Campus de El Carmen s/n, 21071, Huelva, Spain;4. Institute for Environmental Assessment and Water Research (IDÆA), Spanish National Research Council (CSIC), C/Jordi Girona 18-26, Barcelona, Spain;5. Faculdade Meridional IMED, Senador Pinheiro 304, 99070-220, Passo Fundo - RS, Brazil;1. Laboratory of Virology, Institute of Biomedical Sciences, Federal University of Uberlândia, Uberlândia-MG 38405-302, Brazil;2. Institute of Chemistry, University of Campinas-UNICAMP, Campinas-SP 13083-871, Brazil;3. Innovation Center in Salivary Diagnostic and Nanotheranostics, Department of Physiology, Institute of Biomedical Sciences, Federal University of Uberlandia, Minas Gerais, Brazil;4. Department of Fundamental Chemistry, Institute of Chemistry, University of São Paulo, Sao Paulo 05508-000, Brazil;5. Laboratory of Synthesis of Bioinspired Molecules, Institute of Chemistry, Federal University of Uberlândia, Uberlândia-MG 34000-902, Brazil;6. Institute of Biosciences, Humanities and Exact Sciences (Ibilce), São Paulo State University (Unesp), Campus São José do Rio Preto, São José do Rio Preto, São Paulo, Brazil;1. Instituto de Química, Universidade Federal de Uberlândia, Campus Santa Mônica, Uberlândia, MG, Brazil;2. Departamento de Química, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil;3. Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil;4. UNESP – Univ. Estadual Paulista, Institute of Chemistry, Araraquara, SP, Brazil;5. Departamento de Química, Universidade Estadual de Maringá, PR, Brazil;6. Faculdades Associadas de Uberaba, Uberaba, MG, Brazil;7. Laboratório de Biofísica Teórica, Departamento de Física, Instituto de Ciências Exatas, Naturais e Educação, Universidade Federal do Triângulo Mineiro, Uberaba, MG, Brazil;8. Instituto de Química, Universidade Federal Fluminense, Campus Valonguinho, Niterói, RJ, Brazil;9. Departamento de Química, Universidade Federal de Minas Gerais, Campus Pampulha, Belo Horizonte, MG, Brazil;10. Instituto de Ciências Exatas e da Terra, Campus Universitário do Araguaia, Universidade Federal do Mato Grosso, Pontal do Araguaia, MT, Brazil |
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| Abstract: | The reaction of diethylpyrocarbonate (DEPC) with uncoordinated surface histidine residues on metalloproteins at pH 6.5?7.0 can be readily monitored (～30 min) by spectrophotometric changes at ～238 nm (? = 2750 M?1). No reaction is observed if prior modification of histidine by attachment of Ru(II) or Pt(II) has been carried out. In the studies with azurin and cytochrome c only one of two histidines readily undergoes DEPC modification, and likewise only one histidine is Ru modified. Thus the reaction can be used as a quick test for histidine availability, and subsequently as a test as to whether modification has been achieved. The specificity of Pt complexes for histidines is less than that of Ru. In a wider context of histidine availability only one of three histidines in Rieske's protein (N. crassa) is DEPC modified. |
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