scholarly journals Mass Spectrometric Identification of a Novel Factor XIIIa Cross-Linking Site in Fibrinogen

Proteomes ◽  
2021 ◽  
Vol 9 (4) ◽  
pp. 43
Author(s):  
Mariya E. Semkova ◽  
J. Justin Hsuan
Keyword(s):  
Mass Spectrometry ◽  
Protein Function ◽  
Peptide Identification ◽  
Mass Spectrometric ◽  
Factor Xiiia ◽  
Cross Linking ◽  
Spectrometric Analysis ◽  
Peptide Analysis ◽  
Linear Peptide ◽  
Cross Links

Transglutaminases are a class of enzymes that catalyze the formation of a protein:protein cross-link between a lysine and a glutamine residue. These cross-links play important roles in diverse biological processes. Analysis of cross-linking sites in target proteins is required to elucidate their molecular action on target protein function and the molecular specificity of different transglutaminase isozymes. Mass-spectrometry using settings designed for linear peptide analysis and software designed for the analysis of disulfide bridges and chemical cross-links have previously been employed to identify transglutaminase cross-linking sites in proteins. As no control peptide with which to assess and improve the mass spectrometric analysis of TG cross-linked proteins was available, we developed a method for the enzymatic synthesis of a well-defined transglutaminase cross-linked peptide pair that mimics a predicted tryptic digestion product of collagen I. We then used this model peptide to determine optimal score thresholds for correct peptide identification from y- and b-ion series of fragments produced by collision-induced dissociation. We employed these settings in an analysis of fibrinogen cross-linked by the transglutaminase Factor XIIIa. This approach resulted in identification of a novel cross-linked peptide in the gamma subunit. We discuss the difference in behavior of ions derived from different cross-linked peptide sequences and the consequent demand for a more tailored mass spectrometry approach for cross-linked peptide identification compared to that routinely used for linear peptide analysis.

Direct analysis of herbicides by paper spray ionization mass spectrometry

2015 ◽  
Vol 7 (23) ◽  
pp. 9808-9816 ◽  
Author(s):  
Steven L. Reeber ◽  
Sneha Gadi ◽  
Sung-Ben Huang ◽  
Gary L. Glish
Keyword(s):  
Mass Spectrometry ◽  
Environmental Samples ◽  
Direct Analysis ◽  
Mass Spectrometric ◽  
Mass Spectrometric Analysis ◽  
Spectrometric Analysis ◽  
Ionization Mass ◽  
Paper Spray ◽  
Paper Spray Ionization ◽  
Rapid Mass

Paper spray ionization enables the rapid mass spectrometric analysis of environmental samples without the use of chromatography or sample cleanup techniques.

scholarly journals Mass Spectrometric Analysis of Multiple Pertussis Toxins and Toxoids

2010 ◽  
Vol 2010 ◽  
pp. 1-9 ◽  
Author(s):  
Yulanda M. Williamson ◽  
Hercules Moura ◽  
David Schieltz ◽  
Jon Rees ◽  
Adrian R. Woolfitt ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Immune Response ◽  
Mass Spectrometric ◽  
Peptide Sequence ◽  
Spectrometric Analysis ◽  
Vaccine Preventable Disease ◽  
Label Free ◽  
Chemically Modified ◽  
Quantification Method ◽  
Major Virulence Factor

Bordetella pertussis(Bp) is the causative agent of pertussis, a vaccine preventable disease occurring primarily in children. In recent years, there has been increased reporting of pertussis. Current pertussis vaccines are acellular and consist of Bp proteins including the major virulence factor pertussis toxin (Ptx), a 5-subunit exotoxin. Variation in Ptx subunit amino acid (AA) sequence could possibly affect the immune response. A blind comparative mass spectrometric (MS) analysis of commercially available Ptx as well as the chemically modified toxoid (Ptxd) from licensed vaccines was performed to assess peptide sequence and AA coverage variability as well as relative amounts of Ptx subunits. Qualitatively, there are similarities among the various sources based on AA percent coverages and MS/MS fragmentation profiles. Additionally, based on a label-free mass spectrometry-based quantification method there is differential relative abundance of the subunits among the sources.

Extraction and Chromatography-Mass Spectrometric Analysis of the Active Principles from Selected Chinese Herbs and Other Medicinal Plants

2003 ◽  
Vol 31 (06) ◽  
pp. 927-944 ◽  
Author(s):  
Xiaosuo Wang ◽  
Vimal Kapoor ◽  
George A. Smythe
Keyword(s):  
Mass Spectrometry ◽  
Analytical Techniques ◽  
Mass Spectrometric ◽  
Chinese Herbs ◽  
Spectrometric Analysis ◽  
Toxic Substances ◽  
Herbal Extracts ◽  
Active Components ◽  
Herbal Products ◽  
Isolation And Characterization

Medicinal herbs have a long history of use in the practice of traditional Chinese medicine and a substantial body of evidence has, over recent decades, demonstrated a range of important pharmacological properties. Western biomedical researchers are examining not only the efficacy of the traditional herbal products but, through the use of a range of bioassays and analytical techniques, are developing improved methods to isolate and characterize active components. This review briefly describes the different extraction methodologies used in the preparation of herbal extracts and reviews the utility of chromatography-mass spectrometry for the analysis of their active components. In particular, applications of gas or liquid chromatography with mass spectrometry for the isolation and characterization of active components of ginseng are critically assessed. The analysis of toxic substances from herb extracts with mass spectrometric techniques is also discussed along with the potential for mass spectrometric methods to investigate the proteomics of herbal extracts.

scholarly journals Mass spectrometric quantification of the adaptations in the wall proteome of Candida albicans in response to ambient pH

Microbiology ◽  
2011 ◽  
Vol 157 (1) ◽  
pp. 136-146 ◽  
Author(s):  
Grazyna J. Sosinska ◽  
Leo J. de Koning ◽  
Piet W. J. de Groot ◽  
Erik M. M. Manders ◽  
Henk L. Dekker ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Candida Albicans ◽  
Pathogenic Fungus ◽  
Hyphal Growth ◽  
Mass Spectrometric ◽  
Spectrometric Analysis ◽  
Mucosal Surfaces ◽  
Covalently Linked ◽  
Semi Solid

The mucosal layers colonized by the pathogenic fungus Candida albicans differ widely in ambient pH. Because the properties and functions of wall proteins are probably pH dependent, we hypothesized that C. albicans adapts its wall proteome to the external pH. We developed an in vitro system that mimics colonization of mucosal surfaces by growing biomats at pH 7 and 4 on semi-solid agarose containing mucin as the sole nitrogen source. The biomats expanded radially for at least 8 days at a rate of ∼30 μm h−1. At pH 7, hyphal growth predominated and growth was invasive, whereas at pH 4 only yeast and pseudohyphal cells were present and growth was noninvasive. Both qualitative mass spectrometric analysis of the wall proteome by tandem mass spectrometry and relative quantification of individual wall proteins (pH 7/pH 4), using Fourier transform mass spectrometry (FT-MS) and a reference mixture of 15N-labelled yeast and hyphal walls, identified similar sets of >20 covalently linked wall proteins. The adhesion proteins Als1 and Als3, Hyr1, the transglucosidase Phr1, the detoxification enzyme Sod5 and the mammalian transglutaminase substrate Hwp1 (immunological detection) were only present at pH 7, whereas at pH 4 the level of the transglucosidase Phr2 was >35-fold higher than at pH 7. Sixteen out of the 22 proteins identified by FT-MS showed a greater than twofold change. These results demonstrate that ambient pH strongly affects the wall proteome of C. albicans, show that our quantitative approach can give detailed insights into the dynamics of the wall proteome, and point to potential vaccine targets.

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