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A high-temporal resolution technology for dynamic proteomic analysis based on 35S labeling.

Zhang Z, Chen J, Guo F, He L, Wu Y, Zeng C, Xiao X, He D - PLoS ONE (2008)

Bottom Line: The results validated that SiLAD technique, in combination with 2-Dimensional Electrophoresis, provided a highly sensitivity method to illustrate the non-disturbed endogenous proteins dynamic changes with a good temporal resolution and high signal/noise ratio.A significant number of differential proteins can be discovered or re-categorized by this technique.Another unique feature of SiLAD is its capability of quantifying the rate of protein expression, which reflects the cellular physiological turn points more effectively.

View Article: PubMed Central - PubMed

Affiliation: Universities' Confederated Institute of Proteomics, Key laboratory for Cell Proliferation and Regulation Biology Ministry of Education, Beijing Normal University, Beijing, People's Republic of China.

ABSTRACT
As more and more research efforts have been attracted to dynamic or differential proteomics, a method with high temporal resolution and high throughput is required. In present study, a (35)S in vivo Labeling Analysis for Dynamic Proteomics (SiLAD) was designed and tested by analyzing the dynamic proteome changes in the highly synchronized A549 cells, as well as in the rat liver 2/3 partial hepatectomy surgery. The results validated that SiLAD technique, in combination with 2-Dimensional Electrophoresis, provided a highly sensitivity method to illustrate the non-disturbed endogenous proteins dynamic changes with a good temporal resolution and high signal/noise ratio. A significant number of differential proteins can be discovered or re-categorized by this technique. Another unique feature of SiLAD is its capability of quantifying the rate of protein expression, which reflects the cellular physiological turn points more effectively. Finally, the prescribed SiLAD proteome snapshot pattern could be potentially used as an exclusive symbol for characterizing each stage in well regulated biological processes.

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Flow Chart of SiLAD technology.
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pone-0002991-g001: Flow Chart of SiLAD technology.

Mentions: In this study, a novel method to examine the dynamic proteome expression changes based on 35S metabolic pulse labeling was proposed (Figure 1). 2-Dimensional Electrophoresis (2-DE) was employed in this methodology to detect the protein changes, since it is still the best technology to directly visualize the largest number of proteins simultaneously and separately, thus allowing us to detect most of the differential changing proteins before the Mass Spectrometry (MS/MS) identification. Yet, the selected differential proteins still could be identified by MS/MS. The auto-radiography of the 35S short pulse labeled gel provides the information about protein newly expressed without interference from the former existed amount. Using 35S to pulse label several time points in a biological process is, for the first time, allowing us to chase the proteome expression changes at the second order derivate level. The first order derivative level of protein amount change (Sp) in the time course T is Vp, the changing velocity of protein amount. And


A high-temporal resolution technology for dynamic proteomic analysis based on 35S labeling.

Zhang Z, Chen J, Guo F, He L, Wu Y, Zeng C, Xiao X, He D - PLoS ONE (2008)

Flow Chart of SiLAD technology.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2500177&req=5

pone-0002991-g001: Flow Chart of SiLAD technology.
Mentions: In this study, a novel method to examine the dynamic proteome expression changes based on 35S metabolic pulse labeling was proposed (Figure 1). 2-Dimensional Electrophoresis (2-DE) was employed in this methodology to detect the protein changes, since it is still the best technology to directly visualize the largest number of proteins simultaneously and separately, thus allowing us to detect most of the differential changing proteins before the Mass Spectrometry (MS/MS) identification. Yet, the selected differential proteins still could be identified by MS/MS. The auto-radiography of the 35S short pulse labeled gel provides the information about protein newly expressed without interference from the former existed amount. Using 35S to pulse label several time points in a biological process is, for the first time, allowing us to chase the proteome expression changes at the second order derivate level. The first order derivative level of protein amount change (Sp) in the time course T is Vp, the changing velocity of protein amount. And

Bottom Line: The results validated that SiLAD technique, in combination with 2-Dimensional Electrophoresis, provided a highly sensitivity method to illustrate the non-disturbed endogenous proteins dynamic changes with a good temporal resolution and high signal/noise ratio.A significant number of differential proteins can be discovered or re-categorized by this technique.Another unique feature of SiLAD is its capability of quantifying the rate of protein expression, which reflects the cellular physiological turn points more effectively.

View Article: PubMed Central - PubMed

Affiliation: Universities' Confederated Institute of Proteomics, Key laboratory for Cell Proliferation and Regulation Biology Ministry of Education, Beijing Normal University, Beijing, People's Republic of China.

ABSTRACT
As more and more research efforts have been attracted to dynamic or differential proteomics, a method with high temporal resolution and high throughput is required. In present study, a (35)S in vivo Labeling Analysis for Dynamic Proteomics (SiLAD) was designed and tested by analyzing the dynamic proteome changes in the highly synchronized A549 cells, as well as in the rat liver 2/3 partial hepatectomy surgery. The results validated that SiLAD technique, in combination with 2-Dimensional Electrophoresis, provided a highly sensitivity method to illustrate the non-disturbed endogenous proteins dynamic changes with a good temporal resolution and high signal/noise ratio. A significant number of differential proteins can be discovered or re-categorized by this technique. Another unique feature of SiLAD is its capability of quantifying the rate of protein expression, which reflects the cellular physiological turn points more effectively. Finally, the prescribed SiLAD proteome snapshot pattern could be potentially used as an exclusive symbol for characterizing each stage in well regulated biological processes.

Show MeSH