A Review on Innovating the Concept and Practice of Two-Dimensional Gel Electrophoresis in the Analysis of Proteomes at the Proteoform Level

Zhan, Xianquan and Li, Biao and Zhan, Xiaohan and Schlüter, Hartmut and Jungblut, Peter R. and Coorssen, Jens R. (2021) A Review on Innovating the Concept and Practice of Two-Dimensional Gel Electrophoresis in the Analysis of Proteomes at the Proteoform Level. In: Current Advances in Chemistry and Biochemistry Vol. 2. B P International, pp. 135-152. ISBN 978-93-90768-44-8

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Abstract

Two-dimensional gel electrophoresis (2DE) is an important and well-established technical platform enabling extensive top-down proteomic analysis. However, the long-held but now largely outdated conventional concepts of 2DE have clearly impacted its application to in-depth investigations of proteomes at the level of protein species/proteoforms. It is time to popularize a new concept of 2DE for proteomics. With the development and enrichment of the proteome concept, any given “protein” is now recognized to consist of a series of proteoforms. Thus, it is the proteoform, rather than the canonical protein, that is the basic unit of a proteome, and each proteoform has a specific isoelectric point (pI) and relative mass (Mr). Accordingly, using 2DE, each proteoform can routinely be resolved and arrayed according to its different pI and Mr. Each detectable spot contains multiple proteoforms derived from the same gene, as well as from different genes. Proteoforms derived from the same gene are distributed into different spots in a 2DE pattern. High-resolution 2DE is thus actually an initial level of separation to address proteome complexity and is effectively a pre-fractionation method prior to analysis using mass spectrometry (MS). Furthermore, stable isotope-labeled 2DE coupled with high-sensitivity liquid chromatography-tandem MS (LC-MS/MS) has tremendous potential for the large-scale detection, identification, and quantification of the proteoforms that constitute proteomes. Overall, the beauty and power of this coupled 2DE-LC-MS/MS methodology is its inherent capacity to capitalize on the combination of refined top-down and bottom-up analytical approaches. The capacity for genuine deep proteome analysis at the critical proteoform level is now a definite reality.

Item Type: Book Section
Subjects: Library Keep > Biological Science
Depositing User: Unnamed user with email support@librarykeep.com
Date Deposited: 04 Nov 2023 06:29
Last Modified: 04 Nov 2023 06:29
URI: http://archive.jibiology.com/id/eprint/1765

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