Two-dimensional electrophoretic analysis of proteins expressed by normal and cancerous human crypts: Application of mass spectrometry to peptide-mass fingerprinting

1994 ◽  
Vol 15 (1) ◽  
pp. 391-405 ◽  
Author(s):  
Hong Ji ◽  
Robert H. Whitehead ◽  
Gavin E. Reid ◽  
Robert L. Moritz ◽  
Larry D. Ward ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Peptide Mass Fingerprinting ◽  
Electrophoretic Analysis ◽  
Two Dimensional ◽  
Peptide Mass

Gamma Radiation Induced Intestinal Proteomic Modulation in Mice: A Two Dimensional Electrophoretic Analysis

2017 ◽  
Vol 2 (3) ◽  
pp. 327 ◽  
Author(s):  
Sania Bajaj ◽  
Abhinav Singh ◽  
Bhargab Kalita ◽  
M.H. Yashavarddhan ◽  
Rajiv Ranjan ◽  
...  
Keyword(s):  
Statistical Significance ◽  
Peptide Mass Fingerprinting ◽  
Electrophoretic Analysis ◽  
Two Dimensional ◽  
Peptide Mass ◽  
Damage Response ◽  
Radiation Induced ◽  
High Doses ◽  
Ionising Radiations ◽  
Serious Injuries

<p>Exposure to high doses of radiation causes serious injuries in gastrointestinal tract, by affecting biomolecules of the tissue. To demonstrate the modulation of intestinal proteome by ionising radiations, we analysed changes in protein expression in 9 Gy irradiated C57BL/6 mice at 24 h and 72 h by using two dimensional electrophoresis technique. A total of 19 protein spots with statistical significance (fold change&gt;1.5 and P&lt;0.05) were found to be differentially expressed. Of these 07 spots were identified by MALDI-TOF MS and peptide mass fingerprinting techniques which matched with the known proteins documented in the online database. These proteins belong to biological-functional categories like cytoskeleton system, molecular chaperones, DNA damage response, and stress response. These identified radiation induced proteins can help in understanding the mechanisms behind the intestinal injuries and thus can become potential targets for therapeutics and also aid in drug development.</p>

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 Identification and Phenotypic Characterization of Sphingomonas wittichii Strain RW1 by Peptide Mass Fingerprinting Using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

2005 ◽  
Vol 71 (5) ◽  
pp. 2442-2451 ◽  
Author(s):  
Rolf U. Halden ◽  
David R. Colquhoun ◽  
Eric S. Wisniewski
Keyword(s):  
Mass Spectrometry ◽  
Laser Desorption ◽  
Peptide Mass Fingerprinting ◽  
Phenotypic Characterization ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Flight Mass Spectrometry ◽  
Peptide Mass ◽  
Sphingomonas Wittichii

ABSTRACT Mass spectrometry is a potentially attractive means of monitoring the survival and efficacy of bioaugmentation agents, such as the dioxin-mineralizing bacterium Sphingomonas wittichii strain RW1. The biotransformation activity of RW1 phenotypes is determined primarily by the presence and concentration of the dioxin dioxygenase, an enzyme initiating the degradation of both dibenzo-p-dioxin and dibenzofuran (DF). We explored the possibility of identifying and characterizing putative cultures of RW1 by peptide mass fingerprinting (PMF) targeting this characteristic phenotypic biomarker. The proteome from cells of RW1—grown on various media in the presence and absence of DF—was partially purified, tryptically digested, and analyzed using matrix-assisted laser desorption ionization-time of flight mass spectrometry. Mascot online database queries allowed statistically significant identification of RW1 in disrupted, digested cells (P < 0.01 to 0.05) and in digested whole-cell extracts (P < 0.00001 to 0.05) containing hundreds of proteins, as determined by two-dimensional gel electrophoresis. Up to 14 peptide ions of the alpha subunit of the dioxin dioxygenase (43% protein coverage) were detected in individual samples. A minimum of 107 DF-grown cells was required to identify dioxin degradation-enabled phenotypes. The technique hinges on the detection of multiple characteristic peptides of a biomarker that can reveal at once the identity and phenotypic properties of the microbial host expressing the protein. The results demonstrate the power of PMF of minimally processed microbial cultures as a sensitive and specific technique for the positive identification and phenotypic characterization of certain microorganisms used in biotechnology and bioremediation.

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