scholarly journals Red cell proteins. I. Two-dimensional mapping of human erythrocyte lysate proteins

Blood ◽  
1979 ◽  
Vol 53 (6) ◽  
pp. 1121-1132
Author(s):  
JJ Edwards ◽  
NG Anderson ◽  
SL Nance ◽  
NL Anderson
Keyword(s):  
Human Erythrocyte ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Cell Protein ◽  
Red Cell ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Mapping Techniques ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Dimensional Electrophoresis

Human erythrocyte lysate proteins were resolved into over 250 discrete spots by two-dimensional electrophoresis using isoelectric focusing in the first dimension and electrophoresis in the presence of sodium dodecyl sulfate, (SDS) in the second. The overwhelming excess of hemoglobin has made such analyses difficult in the past. However, with the ISO-DALT two-dimensional electrophoresis system, large numbers of red cell proteins can be mapped in the presence of hemoglobin. When hemoglobin and several other major proteins are removed by adsorption to DEAE-cellulose, additional minor components are seen, giving a total of over 275. With the use of purified preparations, the map positions of five cell enzymes or their subunits were determined: pyruvate kinase, catalase, glucose-6-phosphate dehydrogenase, hypoxanthine phosphoribosyltransferase, and carbonic anhydrase. The mapping techniques described complement and extend those traditionally used to find human red cell protein variants.

scholarly journals Red cell proteins. I. Two-dimensional mapping of human erythrocyte lysate proteins

Blood ◽  
1979 ◽  
Vol 53 (6) ◽  
pp. 1121-1132 ◽  
Author(s):  
JJ Edwards ◽  
NG Anderson ◽  
SL Nance ◽  
NL Anderson
Keyword(s):  
Human Erythrocyte ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Cell Protein ◽  
Red Cell ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Mapping Techniques ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Dimensional Electrophoresis

Abstract Human erythrocyte lysate proteins were resolved into over 250 discrete spots by two-dimensional electrophoresis using isoelectric focusing in the first dimension and electrophoresis in the presence of sodium dodecyl sulfate, (SDS) in the second. The overwhelming excess of hemoglobin has made such analyses difficult in the past. However, with the ISO-DALT two-dimensional electrophoresis system, large numbers of red cell proteins can be mapped in the presence of hemoglobin. When hemoglobin and several other major proteins are removed by adsorption to DEAE-cellulose, additional minor components are seen, giving a total of over 275. With the use of purified preparations, the map positions of five cell enzymes or their subunits were determined: pyruvate kinase, catalase, glucose-6-phosphate dehydrogenase, hypoxanthine phosphoribosyltransferase, and carbonic anhydrase. The mapping techniques described complement and extend those traditionally used to find human red cell protein variants.

Evaluation of lipoproteins and apolipoproteins in serum of a Tangier patient by micro-scale two-dimensional electrophoresis.

1987 ◽  
Vol 33 (4) ◽  
pp. 468-472 ◽  
Author(s):  
T Manabe ◽  
S Visvikis ◽  
M F Dumon ◽  
M Clerc ◽  
G Siest
Keyword(s):  
Molecular Mass ◽  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Disease Patient ◽  
High Density Lipoproteins ◽  
Two Dimensional ◽  
High Concentration ◽  
Two Dimensional Electrophoresis ◽  
Micro Scale ◽  
Dimensional Electrophoresis

Abstract We examined lipoproteins and apolipoproteins in serum of a Tangier-disease patient. We used three different techniques of micro-scale two-dimensional electrophoresis: (a) no denaturants; (b) with sodium dodecyl sulfate (SDS) used only in the slab gel electrophoresis; (c) and with urea and a detergent used in isoelectric focusing and with SDS in slab gel electrophoresis. By technique a, an extremely low concentration of high-density lipoproteins (HDL) in the Tangier serum was seen, and lipoproteins that cannot form HDL complexes were detected as multiple spots in the acidic (pl 4 approximately 5) and relatively low apparent molecular mass (20,000 approximately 80,000) region. By technique b, Tangier low-molecular-mass lipoproteins were dissociated into their constituent apolipoproteins, and we observed a higher proportion of apoC-III, together with lower proportions of apoA-I and apoA-II, than in the normal HDL fraction. Technique c showed the total content of apolipoproteins in the whole Tangier serum, as several workers have reported. The presence of low-molecular-mass lipoproteins and a high concentration of apoC-III in this lipoprotein fraction characterized the Tangier serum.

Comparison of yeast cell protein solubilization procedures for two-dimensional electrophoresis

2007 ◽  
pp. 246-249
Author(s):  
Alois Harder ◽  
Robert Wildgruber ◽  
Arek Nawrocki ◽  
Stephen J. Fey ◽  
Peter Mose Larsen ◽  
...  
Keyword(s):  
Yeast Cell ◽  
Cell Protein ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Protein Solubilization ◽  
Dimensional Electrophoresis

Comparison of yeast cell protein solubilization procedures for two-dimensional electrophoresis

1999 ◽  
Vol 20 (4-5) ◽  
pp. 826-829 ◽  
Author(s):  
Alois Harder ◽  
Robert Wildgruber ◽  
Arek Nawrocki ◽  
Stephen J. Fey ◽  
Peter Mose Larsen ◽  
...  
Keyword(s):  
Yeast Cell ◽  
Cell Protein ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Protein Solubilization ◽  
Dimensional Electrophoresis

An attempt to resolve all the various proteins in a single human cell type by two-dimensional electrophoresis: I. Extraction of all cell proteins.

1984 ◽  
Vol 30 (12) ◽  
pp. 2014-2020 ◽  
Author(s):  
J Klose ◽  
E Zeindl
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Human Cell ◽  
Sodium Dodecyl ◽  
Two Dimensional ◽  
Cell Type ◽  
Two Dimensional Electrophoresis ◽  
Extraction Procedures ◽  
Dodecyl Sulfate ◽  
Protein Classes ◽  
Dimensional Electrophoresis

Abstract A concept is presented for estimation of the total number of different proteins in a single human cell type (exemplified here by Hep cells) by use of two-dimensional electrophoresis (2DE). This concept includes three problems, the first, investigated in this study, being the transfer of all protein species of the cells into a sample useful for separation by 2DE. Five different extraction media containing--in various combinations--urea, Nonidet P-40, Zwittergent, mercaptoethanol, dithiothreitol, and sodium dodecyl sulfate were used step by step in three different extraction procedures to extract the cell proteins. The amount of radiolabeled proteins in each extract was measured. Each extract was subjected to 2DE. From the total mass of cell proteins, 99.99% could be extracted in two steps: 96% were extracted with urea/beta-mercaptoethanol solution, the remaining 4% with sodium dodecyl sulfate/urea/beta-mercaptoethanol solution. A special class of proteins assumed to be present in the latter fraction was not detected. Thus this fraction can be omitted from the further analysis of all cell proteins by 2DE. Protein classes that possibly remain undetected by the described extraction procedures are mentioned.

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