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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

Formation of toxin from B. anthracis. Protective antigen is cleaved and oligomerizes, then binds lethal factor to form lethal toxin, edema factor to form edema toxin, or both to form a mixed toxin.
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toxins-07-03497-f001: Formation of toxin from B. anthracis. Protective antigen is cleaved and oligomerizes, then binds lethal factor to form lethal toxin, edema factor to form edema toxin, or both to form a mixed toxin.

Mentions: B. anthracis produces lethal factor (LF), edema factor (EF), and protective antigen (PA) [2]. Protective antigen is an 83 kDa protein which is cleaved into a 63 kDa portion (PA63) which forms heptamers and octamers that binds to cell surface receptors [3,4,5]. The PA63 oligomer also combines with three to four molecules of lethal factor as seen in Figure 1 to form lethal toxin (LTx) or edema factor to form edema toxin (ETx) or may bind both to form a mixed toxin. LF is a zinc-dependent endoprotease which cleaves mitogen activated protein kinase kinase (MAPKK) [6], and EF a calcium and calmodulin-dependent adenylyl cyclase that converts ATP to cyclic AMP [7]. Both toxins work together to cause disruption of the immune system, septicemia, hemorrhage, and shock, which can lead to death [8,9,10].


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

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

Formation of toxin from B. anthracis. Protective antigen is cleaved and oligomerizes, then binds lethal factor to form lethal toxin, edema factor to form edema toxin, or both to form a mixed toxin.
© Copyright Policy
Related In: Results  -  Collection

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

toxins-07-03497-f001: Formation of toxin from B. anthracis. Protective antigen is cleaved and oligomerizes, then binds lethal factor to form lethal toxin, edema factor to form edema toxin, or both to form a mixed toxin.
Mentions: B. anthracis produces lethal factor (LF), edema factor (EF), and protective antigen (PA) [2]. Protective antigen is an 83 kDa protein which is cleaved into a 63 kDa portion (PA63) which forms heptamers and octamers that binds to cell surface receptors [3,4,5]. The PA63 oligomer also combines with three to four molecules of lethal factor as seen in Figure 1 to form lethal toxin (LTx) or edema factor to form edema toxin (ETx) or may bind both to form a mixed toxin. LF is a zinc-dependent endoprotease which cleaves mitogen activated protein kinase kinase (MAPKK) [6], and EF a calcium and calmodulin-dependent adenylyl cyclase that converts ATP to cyclic AMP [7]. Both toxins work together to cause disruption of the immune system, septicemia, hemorrhage, and shock, which can lead to death [8,9,10].

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