title: Nitroproteins in Human Astrocytomas Discovered by Gel Electrophoresis and Tandem Mass Spectrometry.
abstract: Protein tyrosine nitration is involved in the pathogenesis of highly fatal astrocytomas, a type of brain cancer. To understand the molecular mechanisms of astrocytomas and to discover new biomarkers/therapeutic targets, we sought to identify nitroproteins in human astrocytoma tissue. Anti-nitrotyrosine immunoreaction-positive proteins from a high-grade astrocytoma tissue were detected with two-dimensional gel electrophoresis (2DGE)-based nitrotyrosine immunoblots, and identified with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Fifty-seven nitrotyrosine immunopositive protein spots were detected. A total of 870 proteins (nitrated and non-nitrated) in nitrotyrosine-immunopositive 2D gel spots were identified, and 18 nitroproteins and their 20 nitrotyrosine sites were identified with MS/MS analysis. These nitroproteins participate in multiple processes, including drug-resistance, signal transduction, cytoskeleton, transcription and translation, cell proliferation and apoptosis, immune response, phenotypic dedifferentiation, cell migration, and metastasis. Among those nitroproteins that might play a role in astrocytomas was nitro-sorcin, which is involved in drug resistance and metastasis and might play a role in the spread and treatment of an astrocytoma. Semiquantitative immune-based measurements of different sorcin expressions were found among different grades of astrocytomas relative to controls, and a semiquantitative increased nitration level in high-grade astrocytoma relative to control. Nitro-β-tubulin functions in cytoskeleton and cell migration. Semiquantitative immunoreactivity of β-tubulin showed increased expression among different grades of astrocytomas relative to controls and semiquantitatively increased nitration level in high-grade astrocytoma relative to control. Each nitroprotein was rationalized and related to the corresponding functional system to provide new insights into tyrosine nitration and its potential role in the pathogenesis of astrocytoma formation. Graphical Abstract ᅟ.
authors.affiliation: Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Hunan, 410008, People's Republic of China.|Hunan Engineering Laboratory for Structural Biology and Drug Design, Xiangya Hospital, Central South University, Hunan, 410008, People's Republic of China.|State Local Joint Engineering Laboratory for Anticancer Drugs, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, People's Republic of China.
authors.forename: Fang
authors.initials: F
authors.lastname: Peng
authors.affiliation: Molecular Science and Biomedicine Laboratory, State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Biology, Hunan University, Changsha, Hunan, 410018, People's Republic of China.
authors.forename: Jianglin
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Hunan, 410008, People's Republic of China.|Hunan Engineering Laboratory for Structural Biology and Drug Design, Xiangya Hospital, Central South University, Hunan, 410008, People's Republic of China.|State Local Joint Engineering Laboratory for Anticancer Drugs, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, People's Republic of China.
authors.forename: Tianyao
authors.initials: T
authors.lastname: Guo
authors.affiliation: Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Hunan, 410008, People's Republic of China.|Hunan Engineering Laboratory for Structural Biology and Drug Design, Xiangya Hospital, Central South University, Hunan, 410008, People's Republic of China.|State Local Joint Engineering Laboratory for Anticancer Drugs, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, People's Republic of China.|Department of Lung Cancer and Gastroenterology, Hunan Cancer Hospital, Changsha, Hunan, 410013, People's Republic of China.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Yang
authors.affiliation: Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Hunan, 410008, People's Republic of China.|Hunan Engineering Laboratory for Structural Biology and Drug Design, Xiangya Hospital, Central South University, Hunan, 410008, People's Republic of China.|State Local Joint Engineering Laboratory for Anticancer Drugs, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, People's Republic of China.
authors.forename: Maoyu
authors.initials: M
authors.lastname: Li
authors.affiliation: Department of Neurosurgery, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, People's Republic of China.
authors.forename: Shushan
authors.initials: S
authors.lastname: Sang
authors.affiliation: Department of Neurosurgery, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, People's Republic of China.
authors.forename: Xuejun
authors.initials: X
authors.lastname: Li
authors.affiliation: The Charles B. Stout Neuroscience Mass Spectrometry Laboratory, Department of Neurology, College of Medicine, University of Tennessee Health Science Center, Memphis, TN, 38163, USA.
authors.forename: Dominic M
authors.initials: DM
authors.lastname: Desiderio
authors.affiliation: Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Hunan, 410008, People's Republic of China. yjzhan2011@gmail.com.|Hunan Engineering Laboratory for Structural Biology and Drug Design, Xiangya Hospital, Central South University, Hunan, 410008, People's Republic of China. yjzhan2011@gmail.com.|State Local Joint Engineering Laboratory for Anticancer Drugs, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, People's Republic of China. yjzhan2011@gmail.com.|The State Key Laboratory of Medical Genetics, Central South University, Changsha, Hunan, 410008, People's Republic of China. yjzhan2011@gmail.com.
authors.forename: Xianquan
authors.initials: X
authors.lastname: Zhan
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D001254:Astrocytoma / Q000737:chemistry* / Q000473:pathology; D015153:Blotting, Western; D001921:Brain / Q000473:pathology*; D001923:Brain Chemistry; D001932:Brain Neoplasms / Q000737:chemistry* / Q000473:pathology; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D011506:Proteins / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000032:analysis
chemical_list: D011506:Proteins; C002744:3-nitrotyrosine; D014443:Tyrosine
keywords: Astrocytoma; Bioinformatics; Nitroproteome; Nitrotyrosine; Tandem mass spectrometry; Two-dimensional gel electrophoresis; Western blotting