scholarly journals A system for on-line highly sensitive analysis of expression proteome using nanoLC-nanoES/MS/MS and two-dimensional electrophoresis. Towards large-scale analysis and identification of disease-related proteins.

2000 ◽  
Vol 44 (3) ◽  
pp. 185-190 ◽  
Author(s):  
Takao Kawakami ◽  
Fumihiko Usui ◽  
Toshihide Nishimura
Keyword(s):  
Large Scale ◽  
Two Dimensional ◽  
Scale Analysis ◽  
Sensitive Analysis ◽  
Two Dimensional Electrophoresis ◽  
Large Scale Analysis ◽  
Highly Sensitive ◽  
On Line ◽  
Related Proteins ◽  
Dimensional Electrophoresis

scholarly journals Application of two-dimensional electrophoresis and mass spectrometry to screen endometriosis-related proteins

2014 ◽  
Vol 10 (1) ◽  
pp. 95-100
Author(s):  
HANIKEZI TUERXUN ◽  
YANMEI ZHANG ◽  
FEI JI ◽  
AIXINGZI AILI ◽  
XINHUA YANG ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Related Proteins ◽  
Dimensional Electrophoresis

scholarly journals Proteomic analysis of Saccharomyces cerevisiae grown on glucose or glycerol

2021 ◽  
Author(s):  
◽  
Hannah D. Hoang
Keyword(s):  
Mass Spectrometry ◽  
Saccharomyces Cerevisiae ◽  
Large Scale ◽  
Triosephosphate Isomerase ◽  
Lower Amount ◽  
Protein Abundance ◽  
Two Dimensional ◽  
Yeast Saccharomyces Cerevisiae ◽  
Two Dimensional Electrophoresis ◽  
Dimensional Electrophoresis

<p>The goal of this research was to use two-dimensional electrophoresis to examine changes in abundance of enzymes of the glycolytic pathway in the yeast Saccharomyces cerevisiae grown on carbon sources that support either fermentation to ethanol or oxidative metabolism. Large-scale profiling of protein abundances (expression proteomics) often detects changes in protein abundance between physiological states. Such changes in enzyme abundance are often interpreted as evidence of metabolic change although most textbooks emphasise control of enzyme activities not enzyme amount. Two-dimensional difference gel electrophoresis (2DDIGE) was therefore used to examine differences in protein abundance between S. cerevisiae strain BY4741 grown on either glucose (fermentation) or glycerol. Growth on 2% glucose, but not on glycerol, was accompanied by extensive production of ethanol. Doubling times for growth were 2 h 5 min in glucose and 9 h 41 min in glycerol. Conditions for extraction and two-dimensional electrophoresis of proteins were established. One hundred and seventy nine proteins were identified by MALDI mass spectrometry of tryptic digests of protein spots excised from Coomassie stained gels. All of the enzymes for conversion of glucose to ethanol, except for the second enzyme of glycolysis phosphoglucose isomerase, were identified using twodimensional electrophoresis of 100 μg of protein from cells grown on 2% glucose. Identification of proteins excised from the DIGE gels was more challenging, partly because of the lower amount of protein. Eight of the proteins that showed statistically significant differences in abundance (≥ 2-fold, p ≤ 0.01) between glucose and glycerol were identified by mass spectrometry of proteins excised from the 2DDIGE gels, and a further 18 varying proteins were matched to proteins identified from the Coomassie stained gels. Of these total 26 identified or matched proteins, subunits of five of the enzymes for conversion of glucose to ethanol were more abundant from the fermentative cells grown on glucose. The more abundant glycolytic enzymes were phosphofructokinase 2, fructose-1,6-bisphosphate aldolase, triosephosphate isomerase and enolase, plus pyruvate decarboxylase that was required for conversion of the glycolytic product pyruvate to acetaldehyde. The alcohol dehydrogenases Adh1 and Adh4 that convert acetaldehyde to ethanol were detected but did not vary significantly between growth on glucose or glycerol. The results confirmed that in this case changes in abundance of some enzymes were consistent with the altered metabolic output. Future studies should examine whether changes in the abundance and activity of these enzymes are responsible for the differences in metabolism.</p>

scholarly journals Proteomic analysis of Saccharomyces cerevisiae grown on glucose or glycerol

2021 ◽  
Author(s):  
◽  
Hannah D. Hoang
Keyword(s):  
Mass Spectrometry ◽  
Saccharomyces Cerevisiae ◽  
Large Scale ◽  
Triosephosphate Isomerase ◽  
Lower Amount ◽  
Protein Abundance ◽  
Two Dimensional ◽  
Yeast Saccharomyces Cerevisiae ◽  
Two Dimensional Electrophoresis ◽  
Dimensional Electrophoresis

<p>The goal of this research was to use two-dimensional electrophoresis to examine changes in abundance of enzymes of the glycolytic pathway in the yeast Saccharomyces cerevisiae grown on carbon sources that support either fermentation to ethanol or oxidative metabolism. Large-scale profiling of protein abundances (expression proteomics) often detects changes in protein abundance between physiological states. Such changes in enzyme abundance are often interpreted as evidence of metabolic change although most textbooks emphasise control of enzyme activities not enzyme amount. Two-dimensional difference gel electrophoresis (2DDIGE) was therefore used to examine differences in protein abundance between S. cerevisiae strain BY4741 grown on either glucose (fermentation) or glycerol. Growth on 2% glucose, but not on glycerol, was accompanied by extensive production of ethanol. Doubling times for growth were 2 h 5 min in glucose and 9 h 41 min in glycerol. Conditions for extraction and two-dimensional electrophoresis of proteins were established. One hundred and seventy nine proteins were identified by MALDI mass spectrometry of tryptic digests of protein spots excised from Coomassie stained gels. All of the enzymes for conversion of glucose to ethanol, except for the second enzyme of glycolysis phosphoglucose isomerase, were identified using twodimensional electrophoresis of 100 μg of protein from cells grown on 2% glucose. Identification of proteins excised from the DIGE gels was more challenging, partly because of the lower amount of protein. Eight of the proteins that showed statistically significant differences in abundance (≥ 2-fold, p ≤ 0.01) between glucose and glycerol were identified by mass spectrometry of proteins excised from the 2DDIGE gels, and a further 18 varying proteins were matched to proteins identified from the Coomassie stained gels. Of these total 26 identified or matched proteins, subunits of five of the enzymes for conversion of glucose to ethanol were more abundant from the fermentative cells grown on glucose. The more abundant glycolytic enzymes were phosphofructokinase 2, fructose-1,6-bisphosphate aldolase, triosephosphate isomerase and enolase, plus pyruvate decarboxylase that was required for conversion of the glycolytic product pyruvate to acetaldehyde. The alcohol dehydrogenases Adh1 and Adh4 that convert acetaldehyde to ethanol were detected but did not vary significantly between growth on glucose or glycerol. The results confirmed that in this case changes in abundance of some enzymes were consistent with the altered metabolic output. Future studies should examine whether changes in the abundance and activity of these enzymes are responsible for the differences in metabolism.</p>

scholarly journals GENETIC VARIATION IN PROTEINS: COMPARISON OF ONE-DIMENSIONAL AND TWO-DIMENSIONAL GEL ELECTROPHORESIS

Genetics ◽  
1983 ◽  
Vol 104 (2) ◽  
pp. 381-390
Author(s):  
Tracy McLellan ◽  
Giovanna Ferro-Luzzi Ames ◽  
Kishiko Nikaido
Keyword(s):  
Genetic Variation ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
One Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Related Proteins ◽  
Dimensional Electrophoresis ◽  
Denaturation Of Proteins ◽  
New Mutations ◽  
Limited Sensitivity

ABSTRACT Two proteins with known characteristics on one-dimensional gels were studied by two-dimensional electrophoresis to compare the sensitivities of the two methods in detecting genetic variation. Two-dimensional electrophoresis was found to be less sensitive than several types of one-dimensional gels in distinguishing variants of both proteins. Denaturation of proteins in urea in the two-dimensional method makes it possible to distinguish closely related proteins that differ from each other by units of charge. Many more types of variation in protein sequences can be distinguished on one-dimensional gels in the absence of denaturants. The estimates of heterozygosity based on two-dimensional gels are lower than those based on other methods, at least in part, because of the limited types of sequence differences that can be detected on two-dimensional gels. The application of two-dimensional electrophoresis to the measurement of genetic variation and to the detection of new mutations should be made carefully, in view of the limited sensitivity of the method in finding differences in sequence.

From Proteins to Proteomes: Large Scale Protein Identification by Two-Dimensional Electrophoresis and Arnino Acid Analysis

1996 ◽  
Vol 14 (1) ◽  
pp. 61-65 ◽  
Author(s):  
Marc R. Wilkins ◽  
Christian Pasquali ◽  
Ron D. Appel ◽  
Keli Ou ◽  
Olivier Golaz ◽  
...  
Keyword(s):  
Large Scale ◽  
Protein Identification ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Dimensional Electrophoresis
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