Metalloproteomic approach to liver tissue of rats exposed to mercury

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Santiago, Maria Gabriela A. ; Faria, Victor Diego ; Cirineu, Felipe Dalmazzo ; de Lima Queiroz da Silva, Lucas Luan ; de Almeida, Emerson Carlos ; Cavallini, Nubya Goncalves ; Souza Vieira, Jose Cavalcante ; Henrique Fernandes, Ana Angelica ; Braga, Camila Pereira ; Zara, Luis Fabricio ; Rabelo Buzalaf, Marilia Afonso ; Adamec, Jiri ; Padilha, Pedro de Magalhaes

• Document type: Journal article
• Source: Chemosphere; v. 312, p. 9-pg., 2023-01-01.

Abstract

The aims of this study were to identify mercury-associated protein spots in the liver tissue of rats exposed to low concentrations of mercury and to elucidate the physiological and functional aspects of the proteins identified in the protein spots. Therefore, proteomic analysis of the liver tissue of Wistar rats exposed to mercury chloride (4.60 mu g kg(-1) in Hg2+) was performed for thirty days (Hg-30 group) and sixty days (Hg-60 group). The proteomic profile of the liver tissue of the rats was obtained by two-dimensional electrophoresis (2D-PAGE), and the de-terminations of total mercury in the liver tissue, pellets and protein spots were performed by graphite furnace atomic absorption spectrometry (GFAAS). ImageMaster 2D Platinum 7.0 software was used to identify the differentially expressed mercury-associated protein spots, which were then characterized by liquid chromatog-raphy coupled to tandem mass spectrometry (LC-MS/MS). The determinations by GFAAS indicated a total mercury bioaccumulation of 2812% in the Hg-30 group and 3298% in the Hg-60 group and 10 mercury -associated protein spots with a concentration range of 51 +/- 1.0 to 412 +/- 6.00 mg kg(-1) in the 2D PAGE gels from the liver tissue of the Hg-60 group. The LC-MS/MS analyses allowed the identification of 11 metal binding proteins in mercury-associated protein spots that presented fold change with upregulation >1.5, downregulation <-1.7 or that were expressed only in the Hg-60 group. Using the FASTA sequences of the proteins identified in the mercury-associated protein spots, bioinformatics analyses were performed to elucidate the physiological and functional aspects of the metal binding proteins, allowing us to infer that enzymes such as GSTM2 presented greater mercury concentrations and downregulation <-3; Acaa2 and Bhmt, which showed expression only in the Hg-60 group, among others, may act as potential mercury exposure biomarkers. (AU)

• FAPESP's process: 17/09466-3 - Elucidation of physiological and functional aspects of proteins associated with mercury in pirarucu (Arapaima gigas)
• Grantee: Izabela da Cunha Bataglioli
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 19/02538-4 - Effect of mercury exposure frequency in riverside populations from the Amazon region: identification of possible biomarkers
• Grantee: Pedro de Magalhães Padilha
• Support Opportunities: Regular Research Grants
• FAPESP's process: 19/05298-4 - Identification of proteins associated to mercury in hepatic tissue of rats: study of the effect of exposure frequency
• Grantee: Maria Gabriela de Albuquerque Santiago
• Support Opportunities: Scholarships in Brazil - Scientific Initiation