Abstract
In an effort to compare the molecular basis of differential toxic activity of botulinum neurotoxin A (BoNT/A) and BoNT/E, we have analyzed their membrane channel activity by measuring calcein release from liposomes. Both BoNT/A and /E showed a same level of membrane channel activity that was specifically blocked by IgG specific to the neurotoxins. With the use of fluorescein-labeled dextran, we determined that the size of the channel is at least 24.2 Å which is appropriate for the translocation of a protein of 50 kDa (the light chain of BoNT). These findings would suggest that the difference in the toxicity level of the two BoNT serotypes might reflect differences in either endopeptidase activity or their binding to receptor(s).
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Abbreviations
- BoNT:
-
botulinum neurotoxin
- EDTA:
-
ethylenediaminetetraacetate
- ELISA:
-
Enzyme Linked Immunosorbent Assay
- FITC:
-
fluorescein isothiocyante
- HC:
-
heavy chain
- IgG:
-
immunoglobulin G
- LC:
-
light chain
- TeNT:
-
tetanus toxin
References
Agarwal R., Binz T., Swaminathan S. (2005) Biochemistry 44:8291–8302
Ahnert-Hilger G., Bigalke H. (1995) Prog. Neurobiol. 46:83–96
Atassi M.Z., Dolimbek B.Z. (2004) Protein J. 23:39–52
Bade S., Rummel A., Reisinger C., Karnath T., Ahnert-Hilger G., Bigalke H., Binz T. (2004) J. Neurochem. 91:1461–1472
Blaustein R.O., Germann W.J., Finkelstein A., DasGupta B.R. (1987) FEBS Lett. 226:115–120
Cai S., Sarkar H.K., Singh B.R. (1999) Biochemistry 38:6903–6910
Cai, S., Kukreja, R., Shoesmith, S., Chang, T.Z. and Singh, B.R. (2006). Protein J. 25: 455–462.
Call J.E., Cooke P.H., Miller A.J. (1995) J Appl. Bacteriol. 79:257–263
Chen F., Kuziemko G.M., Amersdorfer P., Wong C., Marks J.D., Stevens R.C. (1997) Infect Immun. 65:1626–1630
Day E.P., Ho J.T., Kunze R.K. Jr, Sun S.T. (1977) Biochim Biophys Acta. 470:503–508
DePaiva A., Poulain B., Lawrence G.W., Shone C.C., Tauc L., Dolly J.O. (1993) J. Biol. Chem. 268:20838–20844
Finkelstein A. (1990) J. Physiol. 84:188–190
Fu F.N., Busath D.D., Singh B.R. (2002) Biophys Chem. 99:17–29
Fu F.N., Singh B.R. (1999) J Protein Chem. 18:701–707
Hammond, K., Caputo, G. A., and London, E. (2002). Biochemistry 41: 3243–3253
Hoch D.H., Romero-Mira M., Ehrlich B.E., Finkelstein A., DasGupta B.R., Simpson L.L. (1985) Proc. Natl. Acad. Sci. USA 82:1692–1696
Kamata Y., Kozaki S. (1994) Biochem. Biophys. Res. Commun. 205:751–757
Keller J.E., Cai F., Neale E.A. (2004) Biochemistry 43:526–532
Koriazova L.K., Montal M. (2003) Nat. Struct. Biol. 10:13–18
Kurtzhals P., Larsen C., Johansen M. (1989) J. Chromatogr. 491:117–127
Lalli G., Bohnert S., Deinhardt K., Verastegui C., Schiavo G. (2003) Trends Microbiol. 11:431–437
Lebeda F.J., Singh B.R. (1999) Toxin Rev. 18:45–76
Li L., Binz T., Niemann H., Singh B.R. (2000) Biochemistry 39:2399–2405
Li L., Singh B.R. (1998) J. Nat.Toxins 7:215–226
Li L., Singh B.R. (1999) Toxin Rev. 18:95–112
Li L., Singh B.R. (2000) Biochemistry 39:6466–6674
Menestrina G., Schiavo G., Montecucco C. (1994) Molec. Aspects Med. 15:79–193
Montal M.S., Blewitt R., Tomich J.M., Montal M. (1991) FEBS Lett. 313:12–18
Ogert R.A., Brown J.E., Singh B.R., Shriver-Lake L.C., Ligler F.S. (1992) Anal. Biochem. 205:306–312
Oh K.J., Senzel L., Collier R.J., Finkelstein A. (1999) Proc. Natl. Acad. Sci. USA 96:8467–8470
Oku N., Kendall D.A., MacDonald R.C. (1982) Biochim. Biophys. Acta 691:332–340
Ratts R., Zeng H., Berg E.A., McComb M.E., Castello C.E., VanderSpek J.C., Murphy J.R. (2003) J. Cell Biol. 160:1139–1150
Rummel A., Mahrhold S., Bigalke H., Binz T. (2004) Mol. Microbiol. 51:631–643
Sakaguchi G. (1982) Pharmacol. Ther. 19:165–194
Sauer H., Pratsch L., Tschopp J., Bhakdi S., Peters R. (1991) Biochim. Biophys. Acta. 1063:137–146
Schantz E.J., Johnson E.A. (1992) Microbiol. Rev. 56:80–99
Simpson L.L. (1986) Annu. Rev. Pharmacol. Toxicol. 26:427–453
Singh B.R. (2000) Nat. Struct. Biol. 7:617–619
Singh B.R., Foley J., Lafontaine C. (1995) J. Protein Chem. 14:7–18
Szoka F. Jr., Papahadjopoulos D. (1978) Proc. Natl. Acad. Sci., USA 75:4194–4198
Acknowledgements
This work was in part supported by a DoD/Army Contract No. W911NF-06–1-0095, and by National Institutes of Health through New England Center of Excellence for Biodefense (AI057159-01).
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Parikh, S., Singh, B.R. Comparative Membrane Channel Size and Activity of Botulinum Neurotoxins A and E. Protein J 26, 19–28 (2007). https://doi.org/10.1007/s10930-006-9040-5
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DOI: https://doi.org/10.1007/s10930-006-9040-5