Synthesis of an antheridiol-inducible polypeptide during sexual morphogenesis of Achlya ambisexualis E87

1985 ◽  
Vol 63 (5) ◽  
pp. 355-365 ◽  
Author(s):  
J. Stephen Horton ◽  
Paul A. Horgen
Keyword(s):  
Gel Electrophoresis ◽  
Transcriptional Control ◽  
Polyacrylamide Gel Electrophoresis ◽  
Casein Hydrolysate ◽  
Electrophoretic Analysis ◽  
Two Dimensional ◽  
Qualitative Synthesis ◽  
Two Dimensional Gel Electrophoresis ◽  
Achlya Ambisexualis

Changes in in vivo polypeptide synthetic patterns during sexual morphogenesis of Achlya ambisexualis E87 were investigated using polyacrylamide gel electrophoresis. Autoradiography of [35S]methionine-labelled polypeptides revealed the apparent qualitative synthesis of a 64 000 dalton polypeptide, commencing 30–60 min after antheridiol addition and continuing for at least 6 h. Synthesis of this pheromone-inducible polypeptide was detected throughout male sexual morphogenesis in still culture matings of the male strain E87 and the female strain 734. Two other species of Achlya were also found to synthesize the 64 000 dalton polypeptide in response to antheridiol. The inducible polypeptide did not appear to be associated strictly with a branching response, as no detectable synthesis occurred when asexual branching was induced by the exogenous addition of either casein hydrolysate or lactalbumin hydrolysate. Two-dimensional gel electrophoretic analysis revealed quantitative changes in the synthesis of many other polypeptides as a result of antheridiol action. In addition, both equilibrium and nonequilibrium two-dimensional gel electrophoresis showed the inducible polypeptide to be basic, demonstrating an isoelectric point of more than 8. Studies using the RNA synthetic inhibitor actinomycin D suggested a transcriptional control of the inducible polypeptide.

scholarly journals Clustering of Clinical Strains ofHelicobacter pylori Analyzed by Two-Dimensional Gel Electrophoresis

2000 ◽  
Vol 7 (2) ◽  
pp. 301-306 ◽  
Author(s):  
Helena Enroth ◽  
Thomas Åkerlund ◽  
Anna Sillén ◽  
Lars Engstrand
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Two Dimensional ◽  
Strain Variation ◽  
Disease Group ◽  
Two Dimensional Gel Electrophoresis ◽  
Specific Proteins ◽  
Patient Groups ◽  
H Pylori ◽  
Disease Specific

ABSTRACT Strain variations of Helicobacter pylori have been tested by numerous methods and compared among different patient groups. The aim of this study was to investigate whether H. pyloriexpresses disease-specific proteins that can be detected by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). H. pylori strains isolated from duodenal ulcer, gastric cancer, and gastritis patients were analyzed. Extensive variation in spot patterns was observed between the strains, but a dendrogram analysis revealed that some strains within each disease group clustered together. Eight proteins were sequenced and found in the H. pylori genome sequence. 2-D PAGE is a useful method for studies of protein expression and for highlighting the extensive strain variation that H. pylori exhibits.

A New Method for Proteome Screening with Two-Dimensional Urea-Sds Polyacrylamide Gel Electrophoresis

2014 ◽  
Vol 1 (1) ◽  
Author(s):  
Areeba Ahmad ◽  
Riaz Ahmad
Keyword(s):  
Gel Electrophoresis ◽  
Protein Separation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cost Effective ◽  
Polyacrylamide Gel ◽  
Present System ◽  
Liver Protein ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis

AbstractTwo-dimensional gel electrophoresis (2DE) separating proteins on the basis of their pI and molecular mass remain the best available technique for protein separation and characterization to date. But due to several limitations, including streak formation in IEF gels, partial solubility of proteins, expensive running conditions and relatively longer time taken, a simple urea-SDS-2D polyacrylamide gel electrophoresis (US2DE) is described here. The system is reasonably sensitive, cost effective with good reproducibility. The method described in this paper employs a chaotropic agent, urea, in the first dimension and sodium dodecyl sulphate (SDS), like conventional system, in the second dimension with an addition of polyacrylamide to screen the liver proteome of healthy and chemically induced fibrotic rats. The system separates the protein on the basis of chargeto- mass ratio and clearly demonstrates differential expression in the liver protein repertoire of healthy and fibrotic rats. Moreover, the present system, like other 2D electrophoretic procedures revealed at least 22 novel spots in the investigated tissues. The technique may be utilized for comprehensive proteome screening of any biological sample and would provide an overview to narrow down the candidate proteins or biomarkers.

scholarly journals Altered Protein Expression of Streptococcus oralis Cultured at Low pH Revealed by Two-Dimensional Gel Electrophoresis

2001 ◽  
Vol 67 (8) ◽  
pp. 3396-3405 ◽  
Author(s):  
Joanna C. Wilkins ◽  
Karen A. Homer ◽  
David Beighton
Keyword(s):  
Gel Electrophoresis ◽  
Peptide Mass Fingerprinting ◽  
Low Ph ◽  
Ionization Mass ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Altered Expression ◽  
Streptococcus Oralis ◽  
Peptide Mass

ABSTRACT Streptococcus oralis is the predominant aciduric nonmutans streptococcus isolated from the human dentition, but the role of this organism in the initiation and progression of dental caries has yet to be established. To identify proteins that are differentially expressed by S. oralis growing under conditions of low pH, soluble cellular proteins extracted from bacteria grown in batch culture at pH 5.2 or 7.0 were analyzed by two-dimensional (2-D) gel electrophoresis. Thirty-nine proteins had altered expression at low pH; these were excised, digested with trypsin using an in-gel protocol, and further analyzed by peptide mass fingerprinting using matrix-assisted laser desorption ionization mass spectrometry. The resulting fingerprints were compared with the genomic database forStreptococcus pneumoniae, an organism that is phylogenetically closely related to S. oralis, and putative functions for the majority of these proteins were determined on the basis of functional homology. Twenty-eight proteins were up-regulated following growth at pH 5.2; these included enzymes of the glycolytic pathway (glyceraldehyde-3-phosphate dehydrogenase and lactate dehydrogenase), the polypeptide chains comprising ATP synthase, and proteins that are considered to play a role in the general stress response of bacteria, including the 60-kDa chaperone, Hsp33, and superoxide dismutase, and three distinct ABC transporters. These data identify, for the first time, gene products that may be important in the survival and proliferation of nonmutans aciduric S. oralis under conditions of low pH that are likely to be encountered by this organism in vivo.

scholarly journals Variations in the Methionine-rich Protein Composition of the Genus Arachis1

1984 ◽  
Vol 11 (1) ◽  
pp. 1-3 ◽  
Author(s):  
Sheikh M. Basha ◽  
Sunil K. Pancholy
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Protein Composition ◽  
Electrophoretic Analysis ◽  
Two Dimensional ◽  
Weight Group ◽  
Two Dimensional Gel Electrophoresis ◽  
One Dimensional ◽  
Molecular Weights ◽  
Isoelectric Points

Abstract Methionine-rich proteins (MRP) from seeds of different species of the Genus Arachis were isolated and analyzed by gel electrophoresis to detect possible compositional differences. One-dimensional gel electrophoretic analysis showed presence of quantitative and qualitative variations among the MRP-fractions. Following two-dimensional gel electrophoresis, the MRP-fractions were found to contain three groups of polypeptides with apparent molecular weights of approximately 21,000; 19,000 and 16,000, and isoelectric points between 5.1 and 5.8. Within each molecular weight group the number of polypeptides varied between 1 and 3.

scholarly journals Two-dimensional gel electrophoretic analysis of the MT3 and DR4 molecules from the different D-typed cells.

1984 ◽  
Vol 160 (3) ◽  
pp. 751-758 ◽  
Author(s):  
M Suzuki ◽  
T Yabe ◽  
M Satake ◽  
T Juji ◽  
H Hamaguchi
Keyword(s):  
Light Chain ◽  
Gel Electrophoresis ◽  
Electrophoretic Analysis ◽  
Structural Heterogeneity ◽  
Class Ii ◽  
Light Chains ◽  
Two Dimensional ◽  
Structural Polymorphism ◽  
Two Dimensional Gel Electrophoresis

This report demonstrates directly, using two-dimensional gel electrophoresis and alloantisera, the following: (a) The DR4 light chains show a structural polymorphism among the Dw4, DKT2, and DYT cells. (b) Most of the class II light chains consist of the DR light chain. (c) The MT3 molecule is distinct from the DR4 molecule in the Dw4, DKT2, and DYT cells. (d) The MT3 molecule does not show any structural heterogeneity among the Dw4, DKT2, and DYT cells. These results suggest that the dissection of the D specificity among Dw4, DKT2, and DYT is mainly caused by the differences of the DR4 molecules.

In vivo distribution of proteins within a single cell.

1982 ◽  
Vol 28 (4) ◽  
pp. 1011-1014 ◽  
Author(s):  
C F Austerberry ◽  
P L Paine
Keyword(s):  
Xenopus Laevis ◽  
Single Cell ◽  
Gel Electrophoresis ◽  
Living Cell ◽  
Experimental System ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
In Vivo Distribution ◽  
Nucleocytoplasmic Distribution

Abstract Using the oocyte of Xenopus laevis, we present an experimental system, involving two-dimensional gel electrophoresis, for measuring unambiguously the nucleocytoplasmic distribution of proteins within a living cell.

scholarly journals Protein extraction method of Metroxylon sagu leaf for high resolution two dimensional gel electrophoresis and comparative proteomics

2019 ◽  
Author(s):  
Mehvish Nisar ◽  
Hasnain Hussain
Keyword(s):  
Gel Electrophoresis ◽  
Extraction Method ◽  
Protein Extraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Fractionation Method ◽  
Metroxylon Sagu ◽  
Time Required

Abstract Objective This study aimed to determine the best protein extraction method of Metroxylon sagu for the two-dimensional gel electrophoresis and the comparative analysis. Results To perform good proteome research, the most critical step is to establish a method that gives the best quality of extracted total proteins. To develop an optimized protein extraction protocol for two-dimensional polyacrylamide gel electrophoresis (2-DE) analysis of Metroxylon sagu, five protein extraction protocols were compared; polyethene glycol (PEG) fractionation method, SDS/phenol method, TCA/acetone method, combination SDS/phenol and TCA/acetone and imidazole method. The PEG fractionation method was found to give the most reproducible gels with the highest number of spots and highest protein concentration followed by SDS/Phenol method. The lowest number of spots were observed in Imidazole method. The PEG fractionation method provides improved resolution and reproducibility of two-dimensional polyacrylamide gel electrophoresis (2-DE) and reduces the time required to analyze samples. Partitioning rubisco by polyethene glycol (PEG) fractionation provides clearer detection of low-abundance protein. Hence the result from this study propose PEG fractionation as the effective protein extraction method for 2-DE proteomic studies of Metroxylon sagu.

scholarly journals Initial Proteome Analysis of Model Microorganism Haemophilus influenzae Strain Rd KW20

2003 ◽  
Vol 185 (15) ◽  
pp. 4593-4602 ◽  
Author(s):  
Eugene Kolker ◽  
Samuel Purvine ◽  
Michael Y. Galperin ◽  
Serg Stolyar ◽  
David R. Goodlett ◽  
...  
Keyword(s):  
Haemophilus Influenzae ◽  
Gel Electrophoresis ◽  
Ribosomal Proteins ◽  
Protein Identification ◽  
Proteome Analysis ◽  
Tandem Mass ◽  
Two Dimensional ◽  
High Confidence ◽  
Two Dimensional Gel Electrophoresis

ABSTRACT The proteome of Haemophilus influenzae strain Rd KW20 was analyzed by liquid chromatography (LC) coupled with ion trap tandem mass spectrometry (MS/MS). This approach does not require a gel electrophoresis step and provides a rapidly developed snapshot of the proteome. In order to gain insight into the central metabolism of H. influenzae, cells were grown microaerobically and anaerobically in a rich medium and soluble and membrane proteins of strain Rd KW20 were proteolyzed with trypsin and directly examined by LC-MS/MS. Several different experimental and computational approaches were utilized to optimize the proteome coverage and to ensure statistically valid protein identification. Approximately 25% of all predicted proteins (open reading frames) of H. influenzae strain Rd KW20 were identified with high confidence, as their component peptides were unambiguously assigned to tandem mass spectra. Approximately 80% of the predicted ribosomal proteins were identified with high confidence, compared to the 33% of the predicted ribosomal proteins detected by previous two-dimensional gel electrophoresis studies. The results obtained in this study are generally consistent with those obtained from computational genome analysis, two-dimensional gel electrophoresis, and whole-genome transposon mutagenesis studies. At least 15 genes originally annotated as conserved hypothetical were found to encode expressed proteins. Two more proteins, previously annotated as predicted coding regions, were detected with high confidence; these proteins also have close homologs in related bacteria. The direct proteomics approach to studying protein expression in vivo reported here is a powerful method that is applicable to proteome analysis of any (micro)organism.

Sign in / Sign up

Export Citation Format

Share Document