title: Proteomic and redox-proteomic study on the role of glutathione reductase in human lung cancer cells.
abstract: Glutathione reductase (GR), a cytosolic protein, plays a vital role in maintaining a correct redox status in cells. However, comprehensive investigations of GR-modulated cellular responses, including protein level alteration and redox regulation, have yet to be performed. In this study, we cultured a human lung adenocarcinoma line transfected with empty pLKO.1 vector as a control, CL1-0shControl, and its GR-knockdown derivative, CL1-0shΔGR, to evaluate differential protein level alteration and redox regulation of these two cell lines. We identified 34 spots that exhibited marked changes in intensities, and 13 proteins showing significant changes in thiol reactivity, in response to GR depletion. Several proteins involved in redox regulation, calcium signaling, cytoskeleton regulation, and protein folding showed significant changes in expression, whereas proteins involved in redox regulation, protein folding, and glycolysis displayed changes in thiol reactivity. Interestingly, GR knockdown induces peroxiredoxin-1 overexpression in the air-exposed tissue and high oxygen consuming tissue such as cornea and liver, but not in the low oxygen consuming tissues such as breast and uterine. In summary, we used a comprehensive lung adenocarcinoma based proteomic approach for identifying GR-modulated protein expression alteration and redox modification. Based on our research, this is the first comprehensive proteomic and redox-proteomic analysis used to investigate the role of GR in a mammalian cell model.
authors.affiliation: Department of Medical Sciences and Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Chiao-Yuan
authors.initials: CY
authors.lastname: Fan
authors.forename: Hsiu-Chuan
authors.initials: HC
authors.lastname: Chou
authors.forename: Yi-Wen
authors.initials: YW
authors.lastname: Lo
authors.forename: Yueh-Feng
authors.initials: YF
authors.lastname: Wen
authors.forename: Yi-Chih
authors.initials: YC
authors.lastname: Tsai
authors.forename: Haimei
authors.initials: H
authors.lastname: Huang
authors.forename: Hong-Lin
authors.initials: HL
authors.lastname: Chan
mesh_terms: D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional; D055785:Gene Knockdown Techniques; D005980:Glutathione Reductase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008175:Lung Neoplasms / Q000201:enzymology* / Q000378:metabolism; D010084:Oxidation-Reduction; D020543:Proteome / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D020543:Proteome; D005980:Glutathione Reductase
keywords: DIGE; Glutathione reductase; MALDI-TOF MS; Proteomics; Redox-proteomics