Rapid detection of apoptosis in mammalian cells by using intact cell MALDI mass spectrometry

The Analyst ◽  
2011 ◽  
Vol 136 (24) ◽  
pp. 5181 ◽  
Author(s):  
Hongjuan Dong ◽  
Wei Shen ◽  
Myra Ting Wai Cheung ◽  
Yimin Liang ◽  
Hon Yeung Cheung ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Rapid Detection ◽  
Mammalian Cells ◽  
Intact Cell ◽  
Maldi Mass Spectrometry

An attempt at taxonomical characterization of some Rhizobial species by intact cell MALDI mass spectrometry

2006 ◽  
Vol 23 (2) ◽  
pp. 177-185 ◽  
Author(s):  
R. R. Prabhu ◽  
M. J. Kulkarni ◽  
V. A. Parasharami ◽  
B. Santhakumari ◽  
S. Paranjape
Keyword(s):  
Mass Spectrometry ◽  
Intact Cell ◽  
Maldi Mass Spectrometry ◽  
Rhizobial Species

scholarly journals Interactions of Gliotoxin with Protein Cysteines

2021 ◽  
Author(s):  
◽  
Rana Fathizargaran
Keyword(s):  
Mass Spectrometry ◽  
Creatine Kinase ◽  
Mammalian Cells ◽  
Protein Modification ◽  
Sulfur Oxidation ◽  
Maldi Mass Spectrometry ◽  
Disulfide Bridge ◽  
Covalent Modification ◽  
Oxidation Products ◽  
Affinity Tag

<p>Gliotoxin is a secondary metabolite that is produced by several species of fungi, and is toxic to mammalian cells. It is immunosuppressive, affects antigen presentation by macrophages, and causes apoptosis of some cells. Gliotoxin is an epipolythiodioxopiperazine molecule and contains an internal disulfide bridge that is highly reactive and essential for its toxicity. Suggested mechanisms of action include modification of thiol groups of cysteine residues in target proteins by generating oxidative stress or through covalent modification. The goal of this project was to develop mass spectrometry methods to detect protein modification by gliotoxin. Creatine kinase was used as a model protein. The measured mass of creatine kinase from 45 spectra gave a mean of 42,944 ± 24 which was consistent with the predicted mass of creatine kinase. A tryptic digest of creatine kinase indicated ions consistent with the predicted masses of the four cys-containing peptides including abundant ions at m/z 794, 1130 and 2870 and an ion at low intensity at 4373. The reaction of creatine kinase with gliotoxin showed a time dependent reaction that after 14 h was consistent with formation of a gliotoxin adduct. Reduction of the product with dithiothreitol released creatine kinase. Analysis of the tryptic peptides using MALDI mass spectrometry indicated complex modification of cysteines possibly including formation of a mixed disulfide adduct, intramolecular disulfides of CK, and sulfur oxidation products. Further analysis using the ICAT (isotope-coded affinity tag) method suggested modification of Cys-254 and Cys-283 by gliotoxin. Preliminary experiments examined the effects of gliotoxin on HL-60 cells using ICAT. Proteins of gliotoxin-treated and untreated cells were labeled with Heavy and Light ICAT reagents. Potential ICAT pairs were detected in the mass spectrum as a preliminary search for proteins affected by gliotoxin. The results indicate that ICAT labeling should be an effective strategy for characterization of the protein targets of gliotoxin.</p>

scholarly journals Intact-Cell MALDI-ToF Mass Spectrometry for the Authentication of Drug-Adapted Cancer Cell Lines

Cells ◽  
2019 ◽  
Vol 8 (10) ◽  
pp. 1194 ◽  
Author(s):  
Jane F. Povey ◽  
Emily Saintas ◽  
Adewale V. Aderemi ◽  
Florian Rothweiler ◽  
Richard Zehner ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cell Line ◽  
Cell Lines ◽  
Mammalian Cells ◽  
Intact Cell ◽  
Mass Spectrometry Analysis ◽  
Genetic Origin ◽  
Whole Cell ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof

The use of cell lines in research can be affected by cell line misidentification. Short tandem repeat (STR) analysis is an effective method, and the gold standard, for the identification of the genetic origin of a cell line, but methods that allow the discrimination between cell lines of the same genetic origin are lacking. Here, we use intact cell MALDI-ToF mass spectrometry analysis, routinely used for the identification of bacteria in clinical diagnostic procedures, for the authentication of a set of cell lines consisting of three parental neuroblastoma cell lines (IMR-5, IMR-32 and UKF-NB-3) and eleven drug-adapted sublines. Principal component analysis (PCA) of intact-cell MALDI-ToF mass spectrometry data revealed clear differences between most, but not all, of the investigated cell lines. Mass spectrometry whole-cell fingerprints enabled the separation of IMR-32 and its clonal subline IMR-5. Sublines that had been adapted to closely related drugs, for example, the cisplatin- and oxaliplatin-resistant UKF-NB-3 sublines and the vincristine- and vinblastine-adapted IMR-5 sublines, also displayed clearly distinctive patterns. In conclusion, intact whole-cell MALDI-ToF mass spectrometry has the potential to be further developed into an authentication method for mammalian cells of a common genetic origin.

MALDI Mass Spectrometry for Rapid Detection and Characterization of Biological Threats

2011 ◽  
pp. 211-224 ◽  
Author(s):  
Nathan A. Hagan ◽  
Jeffrey S. Lin ◽  
Miquel D. Antoine ◽  
Timothy J. Cornish ◽  
Rachel S. Quizon ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Rapid Detection ◽  
Maldi Mass Spectrometry

scholarly journals Intact cell MALDI mass spectrometry biotyping for "at-line" monitoring of apoptosis progression in CHO cell cultures

2013 ◽  
Vol 7 (S6) ◽  
Author(s):  
Sebastian Schwamb ◽  
Bogdan Munteanu ◽  
Björn Meyer ◽  
Carsten Hopf ◽  
Mathias Hafner ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cell Cultures ◽  
Intact Cell ◽  
Maldi Mass Spectrometry ◽  
Cho Cell

scholarly journals Interactions of Gliotoxin with Protein Cysteines

2021 ◽  
Author(s):  
◽  
Rana Fathizargaran
Keyword(s):  
Mass Spectrometry ◽  
Creatine Kinase ◽  
Mammalian Cells ◽  
Protein Modification ◽  
Sulfur Oxidation ◽  
Maldi Mass Spectrometry ◽  
Disulfide Bridge ◽  
Covalent Modification ◽  
Oxidation Products ◽  
Affinity Tag

<p>Gliotoxin is a secondary metabolite that is produced by several species of fungi, and is toxic to mammalian cells. It is immunosuppressive, affects antigen presentation by macrophages, and causes apoptosis of some cells. Gliotoxin is an epipolythiodioxopiperazine molecule and contains an internal disulfide bridge that is highly reactive and essential for its toxicity. Suggested mechanisms of action include modification of thiol groups of cysteine residues in target proteins by generating oxidative stress or through covalent modification. The goal of this project was to develop mass spectrometry methods to detect protein modification by gliotoxin. Creatine kinase was used as a model protein. The measured mass of creatine kinase from 45 spectra gave a mean of 42,944 ± 24 which was consistent with the predicted mass of creatine kinase. A tryptic digest of creatine kinase indicated ions consistent with the predicted masses of the four cys-containing peptides including abundant ions at m/z 794, 1130 and 2870 and an ion at low intensity at 4373. The reaction of creatine kinase with gliotoxin showed a time dependent reaction that after 14 h was consistent with formation of a gliotoxin adduct. Reduction of the product with dithiothreitol released creatine kinase. Analysis of the tryptic peptides using MALDI mass spectrometry indicated complex modification of cysteines possibly including formation of a mixed disulfide adduct, intramolecular disulfides of CK, and sulfur oxidation products. Further analysis using the ICAT (isotope-coded affinity tag) method suggested modification of Cys-254 and Cys-283 by gliotoxin. Preliminary experiments examined the effects of gliotoxin on HL-60 cells using ICAT. Proteins of gliotoxin-treated and untreated cells were labeled with Heavy and Light ICAT reagents. Potential ICAT pairs were detected in the mass spectrum as a preliminary search for proteins affected by gliotoxin. The results indicate that ICAT labeling should be an effective strategy for characterization of the protein targets of gliotoxin.</p>

Time-dependent cortisol turnover in tissues using stable isotope tracers and MALDI Mass Spectrometry (MS) sampling

2018 ◽  
Author(s):  
Shazia Khan ◽  
Diego F Cobice ◽  
Dawn EW Livingstone ◽  
C Logan Mackay ◽  
Scott P Webster ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Stable Isotope ◽  
Maldi Mass Spectrometry ◽  
Time Dependent ◽  
Stable Isotope Tracers ◽  
Isotope Tracers
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