Protein Expression in Vibrio parahaemolyticus 690 Subjected to Sublethal Heat and Ethanol Shock Treatments

2008 ◽  
Vol 71 (11) ◽  
pp. 2289-2294 ◽  
Author(s):  
MING-LUN CHIANG ◽  
WEI-LI HO ◽  
ROCH-CHUI YU ◽  
CHENG-CHUN CHOU
Keyword(s):  
Heat Shock ◽  
Vibrio Parahaemolyticus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Test Organism ◽  
Protein Patterns ◽  
One Dimensional ◽  
Sds Page ◽  
Molecular Masses ◽  
Stressed Cells

Cells of Vibrio parahaemolyticus 690 were subjected either to heat shock at 42°C for 45 min or to ethanol shock in the presence of 5% ethanol for 60 min. The protein profiles of the unstressed and stressed V. parahaemolyticus cells were compared. Additionally, the induction of DnaK- and GroEL-like proteins in the unstressed and stressed cells of V. parahaemolyticus was also examined. Analysis with one-dimensional sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) indicated that three proteins with molecular masses of 93, 77, and 58 kDa were induced by both heat shock and ethanol shock. The protein patterns revealed by two-dimensional electrophoresis were more detailed than those revealed by one-dimensional SDS-PAGE. It was found that heat shock and ethanol shock affected the expression of a total of 28 proteins. Among them, four proteins with molecular masses of 94, 32.1, 26.7, and 25.7 kDa were enhanced by both heat shock and ethanol shock. Furthermore, immunoblot analysis showed the presence of a GroEL-like protein with a molecular mass of 61 kDa in the test organism, with the heat-shocked and ethanol-shocked cells producing a GroEL-like protein in a larger quantity than the unstressed cells. However, DnaK-like protein was not detectable in either the unstressed or the stressed cells.

scholarly journals Evaluation of numerical analysis of SDS-PAGE of protein patterns for typingEnterobacter cloacae

1989 ◽  
Vol 103 (2) ◽  
pp. 265-274 ◽  
Author(s):  
M. Costas ◽  
L. L. Sloss ◽  
R. J. Owen ◽  
M. A. Gaston
Keyword(s):  
Numerical Analysis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Phage Typing ◽  
Clinical Isolates ◽  
Protein Patterns ◽  
One Dimensional ◽  
Sds Page ◽  
Typing Methods ◽  
Type Strains

SUMMARYTwenty cultures comprising 13 clinical isolates ofEnterobacter cloacaefrom two hospitals. the type and another reference stain ofE. cloacaeand the type strains of four otherEnterobactersp. and ofEscherichia coli, were characterized by one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDSPAGE) of whole-cell proteins. The protein patterns were highly reproducible and were used as the basis of a numerical analysis which divided the clinical isolates into nine clearly defined protein types. Comparison with established typing methods indicated that the discrimination of SDS-PAGE was similar to that achieved with conventional typing methods and all strain groups recognized by combined sero/phage typing were also found by SDS-PAGE. In addition, protein typing sub-divided a group of four serotype O3 isolates that were difficult to distinguish by phage typing. We conclude that high-resolution SDS-PAGE of proteins provides an effective method of typing isolates ofE. cloacae.

Effect of heat shock on protein synthesis in flax rust uredosporelings

1985 ◽  
Vol 63 (11) ◽  
pp. 2069-2076 ◽  
Author(s):  
Maichael Shaw ◽  
Rosalinda Boasson ◽  
Leroy Scrubb
Keyword(s):  
Heat Shock ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Heat Stroke ◽  
Sodium Dodecyl ◽  
Axenic Culture ◽  
One Dimensional ◽  
Melampsora Lini ◽  
Molecular Masses ◽  
Shock Proteins

In uredosporelings of Melampsora lini (Ehrenb.) Lév. germinated for 3 h, uptake of [35S]methionine and its incorporation into protein were depressed during a 2-h heat shock induced by transfer from 17 ± 0.5 to 31 °C. Spectrophotometric scans of fluorograms, prepared after one-dimensional sodium dodecyl sulphate – polyacrylamide gel electrophoresis of aliquots of protein extracts containing equal numbers of disintegrations per minute (1 dps = 1 Bq) in protein, showed that heat shock induced statistically significant changes in the relative degrees of incorporation of [35S]methionine into 15 polypeptide bands. Increased labelling occurred in seven bands, which appear to be heat shock proteins with apparent molecular masses of 84, 71, 43.5, 30.5, 19.5, 18, and 17 kD. Decreased labelling occurred in eight bands, which appear to be heat stroke proteins with apparent molecular masses of 56, 54, 48, 46, 34, 32, 31.5, and 14 kD. When [35S]methionine was administered during heat shock at 31 °C the same pattern of polypeptide labelling was observed in extracts made immediately without return to 17 °C and in extracts made after uredosporelings were returned to 17 °C for 24 h. Thus label incorporated into each polypeptide during heat shock was retained for at least 24 h after the return to 17 °C. Administration of [35S]methionine at 17 °C after completion of the heat shock showed that the "normal" or "nonshock" pattern of labelling began to be resumed within 2 h after transfer from 31 to 17 °C. The results are discussed in relation to the effect of heat shock in promoting the initiation and growth of mycelial colonies from uredosporelings in axenic culture.

scholarly journals Identification of outbreak-associated and other strains ofClostridium difficileby numerical analysis of SDS-PAGE protein patterns

1994 ◽  
Vol 113 (1) ◽  
pp. 1-12 ◽  
Author(s):  
M. Costas ◽  
B. Holmes ◽  
M. Ganner ◽  
S. L. W. On ◽  
P. N. Hoffman ◽  
...  
Keyword(s):  
Numerical Analysis ◽  
High Resolution ◽  
Clostridium Difficile ◽  
Gel Electrophoresis ◽  
Protein Pattern ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Patterns ◽  
One Dimensional ◽  
Sds Page

SUMMARYSeventy-three cultures ofClostridium difficileisolated both during, and in the period immediately following, an outbreak of infection in a group of three hospitals, were characterized by one-dimensional sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) of whole-cell proteins. Each protein pattern was characterized by the presence of one or two dense bands which were highly reproducible. The protein patterns were used as the basis for a numerical analysis which divided the strains into five phenons (electrophoretic or EP types). The majority, 60 of the 73 cultures, belonged to a single phenon which included strains from both patients and the environment. We conclude that high-resolution SDS–PAGE of proteins provides an effective method for typingC. difficileand therefore for tracing the possible spread of epidemic strains in hospitals and other institutions, thereby allowing a better understanding of the epidemiology of the organism.

scholarly journals  Analysis of whole cell protein profiles of Salmonella serovars isolated from chicken, turkey and sheep faeces by SDS-PAGE

2010 ◽  
Vol 55 (No. 6) ◽  
pp. 259-263 ◽  
Author(s):  
A. Aksakal
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Visual Assessment ◽  
Cell Protein ◽  
Protein Profiles ◽  
Whole Cell ◽  
Sds Page ◽  
Molecular Masses ◽  
Whole Cell Protein ◽  
Salmonella Serovars

This study was carried out to determine the whole cell protein profiles of Salmonella serovars from chicken, turkey and sheep faeces by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A total of 34 Salmonella strains were included in the study, 14 of them were isolated from chicken, 14 from turkey and six from sheep. SDS-PAGE was carried out using 12% (w/v) separating and 4% (w/v) stacking gels. The results showed more than 30 protein bands ranging in size from 97 kDa (kilodaltons) to below 14.4 kDa as determined by visual assessment of their approximate molecular masses. Protein bands of 78.1, 51.2, 41.5, 37.3, 35.1, 33.9, 30.7, 27.6, 25.4, and 24 kDa were detected in all Salmonella serovars. Salmonella strains used in this study were closely related and could not be differentiated depending on the whole cell protein profiles using SDS-PAGE.

scholarly journals Comparative studies of Yersinia pestis outer membrane isolation techniques and their potential use in plague epidemiology

1990 ◽  
Vol 32 (2) ◽  
pp. 78-83 ◽  
Author(s):  
Frederico Guilherme Coutinho Abath ◽  
Luís Carlos de Sousa Ferreira
Keyword(s):  
Yersinia Pestis ◽  
Outer Membrane ◽  
Outer Membrane Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Patterns ◽  
Isolation Techniques ◽  
Sds Page ◽  
Triton X ◽  
Potential Use

In the present study three techniques for obtaining outer membrane enriched fractions from Yersinia pestis were evaluated. The techniques analysed were: differential solubilization of the cytoplasmic membrane with Sarkosyl or Triton X-100, and centrifugation in sucrose density gradients. The sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of outer membrane isolated by the different methods resulted in similar protein patterns. The measurement of NADH-dehydrogenase and succinate dehydrogenase (inner membrane enzymes) indicated that the outer membrane preparations obtained by the three methods were pure enough for analytical studies. In addition, preliminary evidences on the potential use of outer membrane proteins for the identification of geographic variants of Y. pestis wild isolates are presented.

scholarly journals Digestibility of β-lactoglobulin following cross-linking by trametes versicolor laccase and apple polyphenols

2011 ◽  
Vol 76 (6) ◽  
pp. 847-855 ◽  
Author(s):  
Ziyad Tantoush ◽  
Luka Mihajlovic ◽  
Bojana Kravic ◽  
Jana Ognjenovic ◽  
Ratko Jankov ◽  
...  
Keyword(s):  
Nutritional Value ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cross Linking ◽  
Allergenic Potential ◽  
Sds Page ◽  
Molecular Masses ◽  
Quercetin Glycosides ◽  
Phenolic Extract ◽  
Β Lactoglobulin

?-Lactoglobulin (BLG) is an important nutrient of dairy products and an important allergen in cow?s milk allergy. The aim of this study was to investigate the potential of laccase to cross-link BLG in the presence of an apple phenolic extract (APE) and to characterize the obtained products for their digestibility by pepsin and pancreatin. The composition of the apple phenolics used for cross-linking was determined by LC-ESE-MS. The apple phenolic extract contained significant amounts of quercetin glycosides, catechins and chlorogenic acid. The laccase cross-linked BLG in the presence of apple phenolics. The polymerization rendered the protein insoluble in the reaction mixture. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the cross-linking reaction mixture revealed a heterogeneous mixture of high molecular masses (cross-linked BLG), with a fraction of the BLG remaining monomeric. Enzymatic processing of BLG by laccase and apple polyphenols as mediators can decrease the bi-phasal pepsin- pancreatin digestibility of the monomeric and cross-linked protein, thus decreasing its nutritional value. In addition, reduced BLG digestibility can decrease its allergenic potential. Apple polyphenols can find usage in the creation of new, more functional food products, designed to prevent obesity and hypersensitivity-related disorders.

scholarly journals The Oligomeric State of c Rings from Cyanobacterial F-ATP Synthases Varies from 13 to 15

2007 ◽  
Vol 189 (16) ◽  
pp. 5895-5902 ◽  
Author(s):  
Denys Pogoryelov ◽  
Christian Reichen ◽  
Adriana L. Klyszejko ◽  
René Brunisholz ◽  
Daniel J. Muller ◽  
...  
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Environmental Groups ◽  
Oligomeric State ◽  
Sds Page ◽  
C Subunit ◽  
Molecular Masses ◽  
The Individual ◽  
The Masses ◽  
Atp Synthases

ABSTRACT We isolated the c rings of F-ATP synthases from eight cyanobacterial strains belonging to four different taxonomic classes (Chroococcales, Nostocales, Oscillatoriales, and Gloeobacteria). These c rings showed different mobilities on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), probably reflecting their molecular masses. This supposition was validated with the previously characterized c11, c14, and c15 rings, which migrated on SDS-PAGE in proportion to their molecular masses. Hence, the masses of the cyanobacterial c rings can conveniently be deduced from their electrophoretic mobilities and, together with the masses of the c monomers, allow the calculation of the c ring stoichiometries. The method is a simple and fast way to determine stoichiometries of SDS-stable c rings and hence a convenient means to unambiguously determine the ion-to-ATP ratio, a parameter reflecting the bioenergetic efficacy of F-ATP synthases. AFM imaging was used to prove the accuracy of the method and confirmed that the c ring of Synechococcus elongatus SAG 89.79 is a tridecameric oligomer. Despite the high conservation of the c-subunit sequences from cyanobacterial strains from various environmental groups, the stoichiometries of their c rings varied between c13 and c15. This systematic study of the c-ring stoichiometries suggests that variability of c-ring sizes might represent an adaptation of the individual cyanobacterial species to their particular environmental and physiological conditions. Furthermore, the two new examples of c15 rings underline once more that an F1/Fo symmetry mismatch is not an obligatory feature of all F-ATP synthases.

scholarly journals Numerical analysis of SDS–PAGE protein patterns ofSerratia marcescens: a comparison with other typing methods

1990 ◽  
Vol 105 (1) ◽  
pp. 107-117 ◽  
Author(s):  
B. Holmes ◽  
M. Costas ◽  
L. L. Sloss
Keyword(s):  
Numerical Analysis ◽  
Reference Strain ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Clinical Isolates ◽  
Protein Patterns ◽  
Sds Page ◽  
Typing Methods ◽  
Effective Adjunct ◽  
Reference Strains

SUMMARYTwenty-five cultures comprising 18 clinical isolates ofSerratia marcescensfrom two hospitals, the type strain ofS. marcescens, two reference strains ofS. marinorubra, the type or a reference strain of three other Serratia species and a reference strain of undetermined species, were characterized by one-dimensional sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) of whole-cell proteins. The protein patterns were highly reproducible and were used as the basis of a numerical analysis which divided the clinical isolates into eight protein types. Comparison with O-serotyping indicated that the level of discrimination by SDS–PAGE was similar. As with O-serotyping, a secondary scheme, such as phage typing, is necessary to differentiate strains of the same protein type. We conclude that high-resolution SDS–PAGE of proteins provides an effective adjunct to other methods for typing isolates ofS. marcescens.

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