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Mass Spectrometric Detection of Bacterial Protein Toxins and Their Enzymatic Activity.

Kalb SR, Boyer AE, Barr JR - Toxins (Basel) (2015)

Bottom Line: One such example is botulinum neurotoxin (BoNT), a potent neurotoxin produced by C. botulinum.There are seven known serotypes of BoNT, A-G, and many of the serotypes can be further differentiated into toxin variants, which are up to 99.9% identical in some cases.Mass spectrometric proteomic techniques have been established to differentiate the serotype or toxin variant of BoNT produced by varied strains of C. botulinum.

View Article: PubMed Central - PubMed

Affiliation: Centers for Disease Control and Prevention, 4770 Buford Hwy NE, Atlanta, GA 30341, USA. skalb@cdc.gov.

ABSTRACT
Mass spectrometry has recently become a powerful technique for bacterial identification. Mass spectrometry approaches generally rely upon introduction of the bacteria into a matrix-assisted laser-desorption time-of-flight (MALDI-TOF) mass spectrometer with mass spectrometric recognition of proteins specific to that organism that form a reliable fingerprint. With some bacteria, such as Bacillus anthracis and Clostridium botulinum, the health threat posed by these organisms is not the organism itself, but rather the protein toxins produced by the organisms. One such example is botulinum neurotoxin (BoNT), a potent neurotoxin produced by C. botulinum. There are seven known serotypes of BoNT, A-G, and many of the serotypes can be further differentiated into toxin variants, which are up to 99.9% identical in some cases. Mass spectrometric proteomic techniques have been established to differentiate the serotype or toxin variant of BoNT produced by varied strains of C. botulinum. Detection of potent biological toxins requires high analytical sensitivity and mass spectrometry based methods have been developed to determine the enzymatic activity of BoNT and the anthrax lethal toxins produced by B. anthracis. This enzymatic activity, unique for each toxin, is assessed with detection of the toxin-induced cleavage of strategically designed peptide substrates by MALDI-TOF mass spectrometry offering unparalleled specificity. Furthermore, activity assays allow for the assessment of the biological activity of a toxin and its potential health risk. Such methods have become important diagnostics for botulism and anthrax. Here, we review mass spectrometry based methods for the enzymatic activity of BoNT and the anthrax lethal factor toxin.

No MeSH data available.


Related in: MedlinePlus

Graphical representation of the use of proteomics methods to determine the identity of BoNT via deduction of its amino acid sequence.
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toxins-07-03497-f003: Graphical representation of the use of proteomics methods to determine the identity of BoNT via deduction of its amino acid sequence.

Mentions: Mass spectrometry can be used to determine the presence or absence of any protein. The seven known serotypes of BoNT are 64% or less identical at the amino acid sequence level. The toxins from different serotypes contain hundreds of amino acid differences and can be differentiated from each other through determination of their amino acid sequences as seen in Figure 3. The toxins are first digested with an enzyme such as trypsin which breaks the protein into many smaller peptides. The peptides are then fragmented inside the mass spectrometer to produce MS/MS which are specific for the amino acid sequence of the peptide. By knowing the amino acid sequence of a peptide originating from the toxin, the amino acid sequence and hence the identity of the toxin can be determined.


Mass Spectrometric Detection of Bacterial Protein Toxins and Their Enzymatic Activity.

Kalb SR, Boyer AE, Barr JR - Toxins (Basel) (2015)

Graphical representation of the use of proteomics methods to determine the identity of BoNT via deduction of its amino acid sequence.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591662&req=5

toxins-07-03497-f003: Graphical representation of the use of proteomics methods to determine the identity of BoNT via deduction of its amino acid sequence.
Mentions: Mass spectrometry can be used to determine the presence or absence of any protein. The seven known serotypes of BoNT are 64% or less identical at the amino acid sequence level. The toxins from different serotypes contain hundreds of amino acid differences and can be differentiated from each other through determination of their amino acid sequences as seen in Figure 3. The toxins are first digested with an enzyme such as trypsin which breaks the protein into many smaller peptides. The peptides are then fragmented inside the mass spectrometer to produce MS/MS which are specific for the amino acid sequence of the peptide. By knowing the amino acid sequence of a peptide originating from the toxin, the amino acid sequence and hence the identity of the toxin can be determined.

Bottom Line: One such example is botulinum neurotoxin (BoNT), a potent neurotoxin produced by C. botulinum.There are seven known serotypes of BoNT, A-G, and many of the serotypes can be further differentiated into toxin variants, which are up to 99.9% identical in some cases.Mass spectrometric proteomic techniques have been established to differentiate the serotype or toxin variant of BoNT produced by varied strains of C. botulinum.

View Article: PubMed Central - PubMed

Affiliation: Centers for Disease Control and Prevention, 4770 Buford Hwy NE, Atlanta, GA 30341, USA. skalb@cdc.gov.

ABSTRACT
Mass spectrometry has recently become a powerful technique for bacterial identification. Mass spectrometry approaches generally rely upon introduction of the bacteria into a matrix-assisted laser-desorption time-of-flight (MALDI-TOF) mass spectrometer with mass spectrometric recognition of proteins specific to that organism that form a reliable fingerprint. With some bacteria, such as Bacillus anthracis and Clostridium botulinum, the health threat posed by these organisms is not the organism itself, but rather the protein toxins produced by the organisms. One such example is botulinum neurotoxin (BoNT), a potent neurotoxin produced by C. botulinum. There are seven known serotypes of BoNT, A-G, and many of the serotypes can be further differentiated into toxin variants, which are up to 99.9% identical in some cases. Mass spectrometric proteomic techniques have been established to differentiate the serotype or toxin variant of BoNT produced by varied strains of C. botulinum. Detection of potent biological toxins requires high analytical sensitivity and mass spectrometry based methods have been developed to determine the enzymatic activity of BoNT and the anthrax lethal toxins produced by B. anthracis. This enzymatic activity, unique for each toxin, is assessed with detection of the toxin-induced cleavage of strategically designed peptide substrates by MALDI-TOF mass spectrometry offering unparalleled specificity. Furthermore, activity assays allow for the assessment of the biological activity of a toxin and its potential health risk. Such methods have become important diagnostics for botulism and anthrax. Here, we review mass spectrometry based methods for the enzymatic activity of BoNT and the anthrax lethal factor toxin.

No MeSH data available.


Related in: MedlinePlus