two-dimensional-gel-electrophoresis-coupled-with-mass-spectrometry

Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome

Human pituitary adenoma (PA) is a common tumor that occurs in the human pituitary gland in the hypothalamus-pituitary-targeted organ axis systems, and may ...

Central South University

Two-dimensional gel electrophoresis (2DE) in proteomics is traditionally assumed to contain only one or two proteins in each 2DE spot. However, 2DE resolution is being complemented by the rapid development of high sensitivity mass spectrometers. Here we compared MALDI-MS, LC-Q-TOF MS and LC-Orbitrap Velos MS for the identification of proteins within one spot. With LC-Orbitrap Velos MS each Coomassie Blue-stained 2DE spot contained an average of at least 42 and 63 proteins/spot in an analysis of a human glioblastoma proteome and a human pituitary adenoma proteome, respectively, if a single gel spot was analyzed. If a pool of three matched gel spots was analyzed this number further increased up to an average of 230 and 118 proteins/spot for glioblastoma and pituitary adenoma proteome, respectively. Multiple proteins per spot confirm the necessity of isotopic labeling in large-scale quantification of different protein species in a proteome. Furthermore, a protein abundance analysis revealed that most of the identified proteins in each analyzed 2DE spot were low-abundance proteins. Many proteins were present in several of the analyzed spots showing the ability of 2DE-MS to separate at the protein species level. Therefore, 2DE coupled with high-sensitivity LC-MS has a clearly higher sensitivity as expected until now to detect, identify and quantify low abundance proteins in a complex human proteome with an estimated resolution of about 500 000 protein species. This clearly exceeds the resolution power of bottom-up LC-MS investigations.

How many proteins can be identified in a 2DE gel spot within an analysis of a complex human cancer tissue proteome?

Oxidative/nitrative damage is a crucial element among the complex factors that contribute to lung carcinogenesis. Nitric oxide (NO) free radicals, through chemical modifications such as tyrosine nitration, are significantly involved in lung cancinogenesis and metastasis. NO-mediated protein nitration, which is the addition of the nitro group (-NO2) to position 3 of the phenolic ring of a tyrosine residue, is an important molecular event in lung cancer, and has been studied with mass spectrometry. Nitration is involved in multiple biological processes, including signal transduction, protein degradation, energy metabolism, mitochondrial dysfunction, enzyme inactivation, immunogenic response, apoptosis, and cell death. This article reviews the relationship of NO and its derivates and lung cancer, formation and roles of tyrosine nitration in lung cancer, differences of protein nitration between lung cancer and other inflammatory pulmonary diseases, current status of protein nitration and nitroproteomics in lung cancer, and future perspectives to achieve a better understanding of lung carcinogenesis, for biomarker discovery; and for new diagnostic and prognostic monitoring, and therapeutic targets.

Article	Year
How many proteins can be identified in a 2DE gel spot within an analysis of a complex human cancer tissue proteome? Electrophoresis\| PubMed ID: 29205401	2017
Protein tyrosine nitration in lung cancer: current research status and future perspectives. Current medicinal chemistry\| PubMed ID: 29473494	2018

Yuda Huang

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