title: Identification of Arsenic Direct-Binding Proteins in Acute Promyelocytic Leukaemia Cells.
abstract: The identification of arsenic direct-binding proteins is essential for determining the mechanism by which arsenic trioxide achieves its chemotherapeutic effects. At least two cysteines close together in the amino acid sequence are crucial to the binding of arsenic and essential to the identification of arsenic-binding proteins. In the present study, arsenic binding proteins were pulled down with streptavidin and identified using a liquid chromatograph-mass spectrometer (LC-MS/MS). More than 40 arsenic-binding proteins were separated, and redox-related proteins, glutathione S-transferase P1 (GSTP1), heat shock 70 kDa protein 9 (HSPA9) and pyruvate kinase M2 (PKM2), were further studied using binding assays in vitro. Notably, PKM2 has a high affinity for arsenic. In contrast to PKM2, GSTP1and HSPA9 did not combine with arsenic directly in vitro. These observations suggest that arsenic-mediated acute promyelocytic leukaemia (APL) suppressive effects involve PKM2. In summary, we identified several arsenic binding proteins in APL cells and investigated the therapeutic mechanisms of arsenic trioxide for APL. Further investigation into specific signal pathways by which PKM2 mediates APL developments may lead to a better understanding of arsenic effects on APL.
authors.affiliation: Department of Laboratory Medicine, Huashan Hospital, Fudan University, 12 Central Urumqi Road, Shanghai 200040, China. shmuzt@126.com.
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Shanghai Cancer Center and Key Laboratory of Glycoconjugates Research Ministry of Public Health, Fudan University, Shanghai 200032, China. luhaojie@fudan.edu.cn.
authors.forename: Haojie
authors.initials: H
authors.lastname: Lu
authors.affiliation: State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China. drchenzi2@163.com.
authors.forename: Weijun
authors.initials: W
authors.lastname: Li
authors.affiliation: State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China. coryhu@sibs.ac.cn.|Cancer Research Center, SIBS-Xuhui Central Hospital, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China. coryhu@sibs.ac.cn.
authors.forename: Ronggui
authors.initials: R
authors.lastname: Hu
authors.affiliation: Department of Hematology, Huashan Hospital, Fudan University, Shanghai 200040, China. drchenzi@tom.com.
authors.forename: Zi
authors.initials: Z
authors.lastname: Chen
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry* / Q000494:pharmacology; D000077237:Arsenic Trioxide; D001152:Arsenicals / Q000737:chemistry* / Q000494:pharmacology; D001710:Biotin / Q000737:chemistry; D019921:Biotinylation; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification*; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D051549:Glutathione S-Transferase pi / Q000737:chemistry / Q000302:isolation & purification*; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000188:drug therapy / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification*; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D009363:Neoplasm Proteins / Q000737:chemistry / Q000302:isolation & purification*; D010087:Oxides / Q000737:chemistry* / Q000494:pharmacology; D011485:Protein Binding; D019809:Streptavidin / Q000737:chemistry; D053719:Tandem Mass Spectrometry; D013963:Thyroid Hormones / Q000737:chemistry / Q000302:isolation & purification*; D000097810:Thyroid Hormone-Binding Proteins
chemical_list: D000970:Antineoplastic Agents; D000077237:Arsenic Trioxide; D001152:Arsenicals; D001710:Biotin; D002352:Carrier Proteins; D051549:Glutathione S-Transferase pi; D018840:HSP70 Heat-Shock Proteins; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D009363:Neoplasm Proteins; D010087:Oxides; D019809:Streptavidin; D013963:Thyroid Hormones; D000097810:Thyroid Hormone-Binding Proteins; C492001:HSPA9 protein, human; C496556:GSTP1 protein, human
keywords: LC-MS/MS; acute promyelocytic leukaemia; arsenic-binding protein; arsenic-biotin; pyruvate kinase M2