Limits...
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.

Show MeSH

Related in: MedlinePlus

SiLAD profiles are served to characterize the specific state of the cell cycle progress.Curve 1 was from the CBB spot in Figure 3D. Curve 2 was from the spot in Figure 3G PH-I image labeled with downward arrow. Curve 3 was from the same image with Curve 2, but was the spot labeled with upward arrow. The right panels A and B were the defined bar codes for the two time points labeled with spotted line A and B in the left panel. The curve was made by the software Origin6.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2500177&req=5

pone-0002991-g004: SiLAD profiles are served to characterize the specific state of the cell cycle progress.Curve 1 was from the CBB spot in Figure 3D. Curve 2 was from the spot in Figure 3G PH-I image labeled with downward arrow. Curve 3 was from the same image with Curve 2, but was the spot labeled with upward arrow. The right panels A and B were the defined bar codes for the two time points labeled with spotted line A and B in the left panel. The curve was made by the software Origin6.

Mentions: The comparison of the two kinds of time series 2D profiles obtained by the two visualizing methods (CBB and PH-I), provided dynamic curves for a large number of proteins, with not only their existing amounts but also their synthesis activity. A combination pattern containing a couple of (as few as three) chosen differential proteins could serve as exclusive “bar code” for every time point in a well regulated cellular process. Figure 4 shows the specific bar codes for the different moments in A549 model containing the existing amount and synthesis intensity of three spots (one spot from Figure 3D CBB image and two spots from Figure 3G PH-I image). After our carefully compared, as the curve knots in this figure illustrated, each time points have their exclusive bar code derived from the %volumes of these three spots. After appropriate simulate the spot %volume values of the five time points to form these three curves (Figure 4), a bar code can set for any specific moment besides the original five time points set for the measuring. As the examples shown, for the two time points marked by A and B spotted line in the left panel in Figure 4, there are two respective unique bar code patterns shown in the A and B panels in this Figure. In our experiments, there are always more than enough differential spots ready provided for choice to construct efficient bar code, 164 differential spots in A549 cell line model and 124 differential spots in rat liver hepatectomy model. Of course, a careful selection is still essential to optimize the bar code.


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)

SiLAD profiles are served to characterize the specific state of the cell cycle progress.Curve 1 was from the CBB spot in Figure 3D. Curve 2 was from the spot in Figure 3G PH-I image labeled with downward arrow. Curve 3 was from the same image with Curve 2, but was the spot labeled with upward arrow. The right panels A and B were the defined bar codes for the two time points labeled with spotted line A and B in the left panel. The curve was made by the software Origin6.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0002991-g004: SiLAD profiles are served to characterize the specific state of the cell cycle progress.Curve 1 was from the CBB spot in Figure 3D. Curve 2 was from the spot in Figure 3G PH-I image labeled with downward arrow. Curve 3 was from the same image with Curve 2, but was the spot labeled with upward arrow. The right panels A and B were the defined bar codes for the two time points labeled with spotted line A and B in the left panel. The curve was made by the software Origin6.
Mentions: The comparison of the two kinds of time series 2D profiles obtained by the two visualizing methods (CBB and PH-I), provided dynamic curves for a large number of proteins, with not only their existing amounts but also their synthesis activity. A combination pattern containing a couple of (as few as three) chosen differential proteins could serve as exclusive “bar code” for every time point in a well regulated cellular process. Figure 4 shows the specific bar codes for the different moments in A549 model containing the existing amount and synthesis intensity of three spots (one spot from Figure 3D CBB image and two spots from Figure 3G PH-I image). After our carefully compared, as the curve knots in this figure illustrated, each time points have their exclusive bar code derived from the %volumes of these three spots. After appropriate simulate the spot %volume values of the five time points to form these three curves (Figure 4), a bar code can set for any specific moment besides the original five time points set for the measuring. As the examples shown, for the two time points marked by A and B spotted line in the left panel in Figure 4, there are two respective unique bar code patterns shown in the A and B panels in this Figure. In our experiments, there are always more than enough differential spots ready provided for choice to construct efficient bar code, 164 differential spots in A549 cell line model and 124 differential spots in rat liver hepatectomy model. Of course, a careful selection is still essential to optimize the bar code.

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
Related in: MedlinePlus