Some improvements in two-dimensional gel electrophoresis of proteins

Wilson, David L.; Hall, Michael E.; Stone, George C.; Rubin, Robert W.

doi:10.1016/0003-2697(77)90506-1

Cited by 152 publications

(46 citation statements)

References 10 publications

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“…For the next 20 years, O'Farrell's system was the method of choice for separation and analysis of complex protein mixtures, leading us to say that the analysis of proteome expression started in that middle of the 1970s. 2-DE patterns of proteins from cells, tissues or whole organisms were described [14][15][16][17][18][19][20][21][22][23][24]. Turnover of protein precursors, such as for actin and tubulin into isoforms as result of PTM, using pulse-chase technique and 2-DE were demonstrated (examples [25][26][27][28][29][30][31][32]).…”

Section: -De Historical Evolutionmentioning

confidence: 99%

Two‐dimensional gel electrophoresis and mass spectrometry for biomarker discovery

Penque

2009

Proteomics Clinical Apps

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The proteome project, initiated in 1995, was made possible by 2-DE combined with MS. The project main objective was and remains the identification of all proteins expressed by a cell, tissue or organism in a given time and condition. Following this objective, the global profiling of proteins in health versus pathological state by the 2-DE/MS-based proteomic approach has contributed to the elucidation of the basic mechanisms of disease by discovering candidate disease biomarkers and disease targets for new drug development. This review will briefly summarize the historical evolution of 2-DE up to today, and review 2-DE/MS technology and its specific methods of study of immunoresponse (immunoproteomics), PTM of proteins, complex proteinprotein interactions (interactome), the proteome of cell membrane and intracellular proteome turnover in disease biomarker discovery. 2-DE historical evolutionGlobal profiling of proteins in healthy versus pathological states is a strategy to discover biomarkers for early diagnostics, disease progression, treatment response and novel targets for therapeutic drugs development. The idea of making an inventory of all the proteins in a cell, tissue or organism with normal or abnormal physiology, was (re)initiated in the middle of the 1990s with the proteome project [1]. Since then, many technologies to make the analysis of the proteome a reality have been united.The perfect technological 'marriage' that made the proteome project feasible was between high resolution 2-DE for separation of large numbers of proteins and MS for identification and characterization of the proteins displayed on this gel [2]. However, before this happy union occured, science and technology devoted to protein study had a long journey until 2-DE and MS combination was possible (Fig. 1). It started in the last century, in 1937, when Tiselius [3] introduced electrophoresis technique as a modification of Reiner's early concept (1927) [4] to analyse a complex mixture of proteins such as serum proteins. Only 20 years later, in 1959, electrophoresis on a gel support formed by crosslinked polymerization of Cyanogum 41, a commercial name for two organic monomers, acrylamide and the crosslinking agent, N,N1-methylenebisacrylamide, was shown to be useful and was named PAGE by Raymond and Weintraub [5]. The adaptation of polyacrylamide gel to IEF in a stable pH gradient, developed by Svensson in 1962 [6], was possible by the end of the 1960s [7][8][9]. About this time, the 2-D by combining IEF to separate undenatured proteins according to their charge and gradient SDS-PAGE for a second separation, according to their molecular mass, was for the first time introduced by Kenrick and Margolis (1970) [10]. Only in 1975, was IEF in denaturing conditions, as known today, used to follow the 2-D PAGE and it was described in three independent works [11][12][13]. The most powerful demonstraCorrespondence: Dr. Deborah Penque, Laboratório de Proteó-mica, Departamento de Genética, Instituto Nacional de Saúde, Dr. Ricardo Jorge, I.P., A...

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Section: -De Historical Evolutionmentioning

confidence: 99%

Two‐dimensional gel electrophoresis and mass spectrometry for biomarker discovery

Penque

2009

Proteomics Clinical Apps

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“…The positions of individual protein species on the gel were localized using the fluorographic pattern as a guide. Quantification of proteins was carried out by cutting selected spots from the gel and eluting labeled protein from each gel piece in scintillation fluid before counting (35,46). Procedures of data analysis to determine the magnitude of monensin inhibition of individual fast-transported protein species have also been described (35).…”

Section: Analysis Of Fast-transported Proteins By Twodimensional Gel mentioning

confidence: 99%

Evidence that all newly synthesized proteins destined for fast axonal transport pass through the Golgi apparatus.

Hammerschlag¹,

Stone²,

Bolen³

et al. 1982

The Journal of Cell Biology

124

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Effects of the sodium ionophore, monensin, were examined on the passage from neuronal cell body to axon of materials undergoing fast intracellular transport. In vitro exposure of bullfrog dorsal root ganglia to concentrations of drug <1 .0 UM led to a dose-dependent depression in the amount of fast-transported [3H]leucine-or [3H]glycerol-labeled material appearing in the nerve trunk. Incorporation of either precursor was unaffected . Exposure of a desheathed nerve trunk to similar concentrations of monensin, while ganglia were incubated in drug-free medium, had no effect on transport. With [3H]fucose as precursor, fast transport of labeled glycoproteins was depressed to the same extent as with [3H]leucine; synthesis, again, was unaffected . By contrast, with [3 H]galactose as precursor, an apparent reduction in transport of labeled glycoproteins was accounted for by a marked depression in incorporation . The inference from these findings, that monensin acts to block fast transport at the level of the Golgi apparatus, was supported by ultrastructural examination of the drug-treated neurons. An extensive and selective disruption of Golgi saccules was observed, accompanied by an accumulation of clumped smooth membranous cisternae.Quantitative analyses of 48 individual fast-transported protein species, after separation by two-dimensional gel electrophoresis, revealed that monensin depresses all proteins to a similar extent . These results indicate that passage through the Golgi apparatus is an obligatory step in the intracellular routing of materials destined for fast axonal transport.

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“…Among the 40 variable spots in Imajoh (1981), polypeptides 16 and 17 of C might be produced under the possibility (1), (2), or (3). These spots could be identified because multiple spots per allele did not seem to be accompanied with large differences in molecular weight (Wilson et al 1977;Racine and Langley 1980;Garrels 1980). Since polypeptide 17 was common to S, polypeptide 16 would be the multiplied, allelic, or duplicated spot.…”

Section: Index Of Phylogenetic Similaritymentioning

confidence: 99%

“…These 118 spots automatically rejected possibilities (3) and (4) and left possibilities (1) and (2). Conditions to produce multiple spots per allele were either that the samples had been kept in a freezer for more than 2 years or that the samples had been purified (O'Farrell 1975;Wilson et al 1977;Racine and Langley 1980;Garrels 1980). The number of groups of multiplied spots was relatively very small.…”

Section: Index Of Phylogenetic Similaritymentioning

confidence: 99%

Phylogenetic relationships among the Anopheles hyrcanus species group based on the degree of hybrid development and comparison with phylogenies by other methods.

Takai

Kanda

1986

The Japanese journal of genetics

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Phylogenetic relationships among 7 members of the Anopheles hyrcanus species group of East Asia which contains vectors of human parasitic diseases were studied based on the development of interspecific hybrids; they were compared with phylogenetic relationships obtained from morphological and biochemical works. A study of Takai et al. (1984) on postmating reproductive isolation between 7 members of the An, hyrcanus species group demonstrated various degrees of development among hybrids. It is proposed in this study that hybrid development is correlated with phylogenetic relationship. An index of hybrid development was introduced and was applied to classification of the parental members. It was compared with a classification based on the morphological data of Harrison (1972) which were reanalyzed in this study. The data for polypeptide differences disclosed by two-dimensional gel electrophoresis conducted by Imajoh (1981) were reanalyzed to construct a classification, which was compared with the present results.A common clustering among the three classifications was a grouping of An. argyropus, An, nigerrimus, and An. nitidus. Laboratory colonies of this subgroup were difficult in free-mating in cages and preferred to feed on man, whereas colonies of the other species freely mated in cages and preferred to blood-feed on mice, except that An, crawf ordi did not freely mate in cages. Relation of the vector members to the classification is discussed. A revision in the UPGMA method is also presented.

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Some improvements in two-dimensional gel electrophoresis of proteins

Cited by 152 publications

References 10 publications

Two‐dimensional gel electrophoresis and mass spectrometry for biomarker discovery

Two‐dimensional gel electrophoresis and mass spectrometry for biomarker discovery

Evidence that all newly synthesized proteins destined for fast axonal transport pass through the Golgi apparatus.

Phylogenetic relationships among the Anopheles hyrcanus species group based on the degree of hybrid development and comparison with phylogenies by other methods.

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