Search PubMed for publications by Dr Alvin Fox
Fox A., Stewart G.C, Waller L. N., K. F. Fox, Harley W. M. and Price, R.L.
Carbohydrates and glycoproteins of Bacillus anthracis and related bacteria. J.
Microbiol. Meth. 54: 143-152. 2003.
The spore is the form released in a bioterrorism attack. There is a real need
for definition of new targets for Bacillus anthracis that might be incorporated
into emerging biodetection technologies. Particularly of interest are
macromolecules found in B. anthracis that are 1) spore specific 2) readily
accessible on the spore surface and 3) distinct from those present in related
organisms. One of the few methods to identify the spores of B. anthracis is
based on the presence of rhamnose and 3-O-methyl rhamnose as determined by gas
chromatography-mass spectrometry. Related organisms additionally contain
2-O-methyl rhamnose and fucose. Carbohydrates and glycoproteins of the B. cereus
group of organisms and the related B. subilis group are reviewed here. It is
hypothesized that the spore-specific carbohydrate is a component of the newly
described glycoprotein of the exosporium of B. anthracis. Further work to define
the protein and carbohydrate components of the glycoprotein of B. anthracis
could be highly useful in developing new technologies for rapid biodetection.
Waller L., Fox K.F., Fox A. and Price R.L. Ruthenium red staining for
ultra-structural visualization of a glycoprotein layer surrounding the spore of
Bacillus anthracis and B. subtilis. J. Microbiol. Meth. 58: 23-30. 2004.
Ruthenium red is a polycationic stain used to visualize acid polysaccharides on
the outer surface of cells. Ruthenium red staining followed by electron
microscopic analysis was used to demonstrate the presence of an external
glycoprotein layer surrounding the spore of both Bacillus anthracis and Bacillus
subtilis. This layer is less apparent with traditional staining methods used for
electron microscopy. Renografin gradients were used to purify B. subtilis
spores. These purified spores displayed greatly enhanced staining with ruthenium
red, indicating nonspecific binding of renografin, which has a major
carbohydrate constituent, methylglucamine. For B. anthracis, staining with
ruthenium red was sufficiently intense that it was not significantly enhanced by
renografin purification. In addition to demonstrating a previously undiscovered
layer surrounding the spores of B. subtilis, the results help explain a
long-standing controversy as to ultrastructural differences among these
genetically closely related organisms. Ruthenium red staining provides an
important addition to the identification of surface
glycoproteins in studies to define similarities and differences in the
exosporium layers of Bacillus species.
Waller. L.N., Stump M.J., Fox K.F., Harley W.M., Fox A, Stewart G.C. and
Shahgholi
M. Identification of a second collagen-like glycoprotein produced by Bacillus
anthracis and demonstration of associated spore-specific sugars. J. Bacteriol.
187: 4592-4597. 2005.
Certain carbohydrates (rhamnose, 3-O-methyl rhamnose and galactosamine) have
been demonstrated to be present in Bacillus anthracis spores, but absent in
vegetative cells. Others have demonstrated that these spore-specific sugars are
constituents of the glycoprotein, BclA. In the current work, spore extracts were
separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).
A second collagen-like glycoprotein, BclB, was identified for the first time in
B. anthracis. The protein moiety of this glycoprotein was identified by matrix
assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass
spectrometry (MS) and the carbohydrate components by gas chromatography-mass
spectrometry and tandem mass spectrometry (GC-MS and GC-MS-MS, respectively).
Spore-specific sugars were also demonstrated to be components of BclB. |
