Analysis of arsenic binding proteins in HepG2 cells based on a biotinylated phenylarsenite probe
| Autor: | Kai Nan, Man He, Bin Hu, Beibei Chen |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Gel electrophoresis
chemistry.chemical_classification Spectrometry Mass Electrospray Ionization biology Chemistry Electrospray ionization chemistry.chemical_element Hep G2 Cells Biochemistry DNA-binding protein Analytical Chemistry Arsenic Enzyme Biotinylation biology.protein Environmental Chemistry Humans Electrophoresis Polyacrylamide Gel General Transcription Factor IIH Subunit 1 Carrier Proteins Spectroscopy Polymerase |
| Zdroj: | Analytica chimica acta. 1183 |
| ISSN: | 1873-4324 |
| Popis: | For a deep understanding of arsenic's mutagenicity, carcinogenicity and teratogenicity, the elucidation of arsenic binding proteins in organisms is a necessary prerequisite. Herein, a biotinylated phenylarsenite (Bio-PAO(III)) probe was synthesized for in situ binding to arsenic binding proteins in HepG2 cells. The Bio-PAO(III)-arsenic binding proteins complexes were captured by the prepared streptavidin-magnetic beads (SA-MBs) by specific interaction of biotin-SA. After magnetic separation, the arsenic binding proteins in the eluent was separated by sodium dodecyl sulfate-polyacrylamide - gel electrophoresis, and the in-gel tryptic digested protein bands were subjected to capillary high performance liquid chromatography coupled with electrospray ionization mass spectrometry analysis. 32 kinds of arsenic binding proteins were identified in HepG2 cells, which could be divided into three groups, structure proteins, enzymes related with tricarboxylic acid cycle and fatty synthesis and transcriptional regulator. Poly [ADP-ribose] polymerase 1 and general transcription factor IIH subunit 1 were identified to bind with arsenicals, which may affect the process of nucleotide excision repair in HepG2 cells. |
| Databáze: | OpenAIRE |
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