Hemagglutinin binding mediated protection of botulinum neurotoxin from proteolysis.
Type A Clostridium botulinum, the causative agent of the food poisoning botulism disease, secretes botulinum neurotoxins along with seven neurotoxin associated proteins (NAPs). The function of NAPs has been shown to protect the neurotoxin from acidity, heat, and proteolytic attack in the environmental and gastrointestinal tract during the toxicogenesis of the botulism disease. One of the NAPs, purified from type A botulinum neurotoxin complex, showed hemagglutination activity. A direct interaction has been demonstrated between purified NAP, a 33-kDa hemagglutinin or Hn-33, and the neurotoxin by using Sephadex G-200 column chromatography. Furthermore, Hn-33 has complete resistance against proteolytic attack at pH 2.0 as well as at normal physiological pH. We have investigated digestion of the neurotoxin in the presence and absence of Hn-33. The neurotoxin alone has been found to be more susceptible to the enzymatic digestion than neurotoxin with Hn-33. The presence of Hn-33 changes the proteolytic fragmentation pattern of the neurotoxin. It seems that Hn-33 protects the neurotoxin from proteolysis either by structural modification of the neurotoxin or by blocking the protease accessible sites of the neurotoxin.
Department of Chemistry and Biochemistry, University of Massachusetts, Dartmouth 02747-2300, USA.
S K
SK
Sharma
B R
BR
Singh
D019274:Botulinum Toxins, Type A / Q000737:chemistry* / Q000302:isolation & purification; D002846:Chromatography, Affinity; D002918:Chymotrypsin; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D006388:Hemagglutinins / Q000737:chemistry*; D008970:Molecular Weight; D010434:Pepsin A; D010446:Peptide Fragments / Q000737:chemistry; D013381:Subtilisins; D014357:Trypsin
D003470:Culture Media; D006388:Hemagglutinins; D010446:Peptide Fragments; D013381:Subtilisins; D002918:Chymotrypsin; D014357:Trypsin; D010434:Pepsin A; D019274:Botulinum Toxins, Type A