Nitroproteins from a human pituitary adenoma tissue discovered with a nitrotyrosine affinity column and tandem mass spectrometry

• Published in: Analytical biochemistry Vol. 354; no. 2; pp. 279 - 289
• Main Authors: Zhan, Xianquan, Desiderio, Dominic M.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.07.2006
• Subjects: Adenoma - chemistry; Adenoma - genetics; Adult; Amino Acid Sequence; Bioinformatics; Chromatography, Affinity - methods; Chromatography, Affinity - statistics & numerical data; Human nonfunctional pituitary adenoma; Humans; Male; Molecular Sequence Data; Multiprotein Complexes; Neoplasm Proteins - chemistry; Neoplasm Proteins - genetics; Neoplasm Proteins - isolation & purification; Nitrogen - chemistry; Nitroprotein; Nitroproteomics; Nitrotyrosine affinity column; Pituitary Neoplasms - chemistry; Pituitary Neoplasms - genetics; Protein Structure, Tertiary; Proteomics; Sensitivity and Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - statistics & numerical data; Tandem mass spectrometry; Trypsin; Tyrosine - analogs & derivatives; Nitroprotein; Tandem mass spectrometry; Human nonfunctional pituitary adenoma; Nitrotyrosine affinity column; Nitroproteomics; Bioinformatics
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2006.05.024

The aim of this study was to characterize endogenous nitroproteins, and those proteins that interact with nitroproteins, in a human pituitary nonfunctional adenoma so as to clarify the role of protein nitration in adenomas. A nitrotyrosine affinity column (NTAC) was used to preferentially enrich and isolate endogenous nitroproteins and nitroprotein–protein complexes from a tissue homogenate that was prepared from a human pituitary nonfunctional pituitary adenoma. The preferentially enriched endogenous nitroproteins and nitroprotein–protein complexes were subjected to trypsin digestion, desalination, and tandem mass spectrometry analysis. Nine nitroproteins (Rho-GTPase-activing protein 5, leukocyte immunoglobulin-like receptor subfamily A member 4 precursor, zinc finger protein 432, cAMP-dependent protein kinase type I-beta regulatory subunit, sphingosine-1-phosphate lyase 1, centaurin beta 1, proteasome subunit alpha type 2, interleukin 1 family member 6, and rhophilin 2) and three proteins (interleukin 1 receptor-associated kinase-like 2, glutamate receptor-interacting protein 2, and ubiquitin) that interacted with nitroproteins were discovered. The nitration site of each nitroprotein was located onto the functional domain where nitration occurred, and each nitroprotein was related to a corresponding functional system. Those data indicate that protein nitration might be an important molecular event in the formation of a human pituitary nonfunctional adenoma.