Abstract
The crystal structures of a number of neurotoxins are now available and reveal that all botulinum neurotoxins (BoNTs) have similar structures in general. However, there are differences. These variations and their relation to functional differences will be reviewed. BoNTs A, B, and E have similar structural domains responsible for specific functions in toxicity but have different domain organization. This leads to the difference in speed of onset of toxic effect and its efficacy. Individual domains of botulinum toxins also exhibit differences and these can be correlated to their functional differences. Structural information is also being used in developing countermeasures for botulism. The strategies and their results are discussed.
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References
Gill DM (1982) Bacterial toxins: a table of lethal amounts. Microbiol Rev 46:86–94
Schiavo G, Matteoli M, Montecucco C (2000) Neurotoxins affecting neuroexocytosis. Physiol Rev 80:717–766
Singh BR, Gimenez JA, DasGupta BR (1991) Comparative molecular topography of botulinum neurotoxins from Clostridium butyricum and Clostridium botulinum type E. Biochim Biophys Acta 1077:119–126
Bhidayasiri R, Truong DD (2005) Expanding use of botulinum toxin. J Neuro Sci 235:1–9
Bhidayasiri R, Truong DD (2008) Evidence for effectiveness of botulinum toxin for hyperhidrosis. J Neural Transm 115:641–645
Caya JG, Agni R, Miller JE (2004) Clostridium botulinum and the clinical laboratorian: a detailed review of botulism, including biological warfare ramifications of botulinum toxin. Arch Pathol Med 128:653–662
Cheng CM, Chen JS, Patel RP (2006) Unlabeled uses of botulinum toxins: a review, part 1. Am J Health Syst Pharm 63:145–152
Cheng CM, Chen JS, Patel RP (2006) Unlabeled uses of botulinum toxins: a review, part 2. Am J Health Syst Pharm 63:225–232
Foster KA (2004) The analgesic potential of clostridial neurotoxin derivatives. Expert Opin Investig Drugs 13:1437–1443
Foster KA (2009) Engineered toxins: new therapeutics. Toxicon 54:587–592
Foster KA, Bigalke H, Aoki KR (2006) Botulinum neurotoxin—from laboratory to bedside. Neurotox Res 9:133–140.
Karalewitz AP, Kroken AR, Fu Z, Baldwin MR, Kim JJ, Barbieri JT (2010) Identification of a unique ganglioside binding loop within botulinum neurotoxins C and D-SA. Biochemistry 49:8117–8126.
Montecucco C, Papini E, Schiavo G (1994) Bacterial protein toxins penetrate cells via a four-step mechanism. FEBS Lett 346:92–98
Kumaran D, Eswaramoorthy S, Furey W, Navaza J, Sax M, Swaminathan S (2009) Domain organization in Clostridium botulinum neurotoxin type E is unique: Its implication in faster translocation. J Mol Biol 386:233–245
Lacy DB, Tepp W, Cohen AC, DasGupta BR, Stevens, RC (1998) Crystal structure of botulinum neurotoxin type A and implications for toxicity. Nat Struct Biol 5:898–902
Swaminathan S, Eswaramoorthy S (2000) Structural analysis of the catalytic and binding sites of Clostridium botulinum neurotoxin B. Nat Struct Biol 7:693–699
Lacy DB, Stevens RC (1999) Sequence homology and structural analysis of clostridial neurotoxins. J Mol Biol 291:1091–1104
Umland TC, Wingert LM, Swaminathan S, Furey WF, Schmidt JJ, Sax M (1997) Structure of the receptor binding fragment Hc of tetanus neurotoxin. Nat Struct Biol 4:788–792
Murzin AG, Lesk AM, Chothia C (1992) beta-Trefoil fold. Patterns of structure and sequence in the Kunitz inhibitors interleukins-1 beta and 1 alpha and fibroblast growth factors. J Mol Biol 223:531–543
Fischer A, Garcia-Rodriguez C, Geren I, Lou J, Marks JD, Nakagawa T, Montal M (2008) Molecular architecture of botulinum neurotoxin E revealed by single particle electron microscopy. J Biol Chem 283:3997–4003
Chai Q, Arndt JW, Dong M, Tepp WH, Johnson EA, Chapmann ER, Stevens RC (2006) Structural basis of cell surface receptor recognition by botulinum neurotoxin B. Nature 444:1019–1020
Emsley P, Fotinou C, Black I, Fairweather NF, Charles IG, Watts C, Hewitt E, Isaacs NW (2000) The structures of the H(C) fragment of tetanus toxin with carbohydrate subunit complexes provide insight into ganglioside binding. J Biol Chem 275:8889–8894
Fotinou C, Emsley P, Black I, Ando H, Ishida H, Kiso M, Sinha KA, Fairweather NF, Isaacs NW (2001) The crystal structure of tetanus toxin Hc fragment complexed with a synthetic Gt1b analogue suggests cross-linking between ganglioside receptors and the toxin. J Biol Chem 276:32274–32281
Jayaraman S, Eswaramoorthy S, Ahmed SA, Smith LA, Swaminathan S (2005) N-terminal helix reorients in recombinant C-fragment of Clostridium botulinum type B. Biochem Biophys Res Commun 330:97–103
Jayaraman S, Eswaramoorthy S, Kumaran D, Swaminathan S (2005) Common binding site for disialyllactose and tri-peptide in C-fragment of tetanus neurotoxin. Proteins 61:288–295
Jin R, Rummel A, Binz T, Brunger AT (2006) Botulinum neurotoxin B recognizes its protein receptor with high affinity and specificity. Nature 444:1092–1095
Strotmeier J, Lee K, Volker AK, Mahrhold S, Zong Y, Zeiser J, Zhou J, Pich A, Bigalke H, Binz T, Rummel A, Jin R (2010) Botulinum neurotoxin serotype D attacks neurons via two carbohydrate-binding sites in a ganglioside-dependent manner. Biochem J 431:207–216
Fu Z, Chen C, Barbieri JT, Kim JJ, Baldwin MR (2009) Glycosylated SV2 and gangliosides as dual receptors for botulinum neurotoxin serotype F. Biochemistry 48:5631–5641
Stenmark P, Dupuy J, Imamura A, Kiso M, Stevens RC (2008) Crystal structure of botulinum neurotoxin type A in complex with the cell surface co-receptor GT1b-insight into the toxin-neuron interaction. PLoS Pathog 4:e1000129
Stenmark P, Dong M, Dupuy J, Chapman ER, Stevens RC (2010) Crystal structure of the botulinum neurotoxin type G binding domain: insight into cell surface binding. J Mol Biol 397:1287–1297
Muraro L, Tosatto S, Motterlini L, Rossetto O, Montecucco C (2009) The N-terminal half of the receptor domain of botulinum neurotoxin A binds to microdomains of the plasma membrane. Biochem Biophys Res Commun 380:76–80
Binz T, Rummel A (2009) Cell entry strategy of clostridial neurotoxins. J Neurochem 109:1584–1595
Kitamura M, Takamiya K, Aizawa S, Furukawa K, Furukawa K (1999) Gangliosides are the binding substances in neural cells for tetanus and botulinum toxins in mice. Biochemica Biophysica Acta 1441:1–3
Montecucco C (1986) How do tetanus and botulinum toxins bind to neuronal membranes? Trends Biochem Sci 11:314–317
Bigalke H, Muller H, Dreyer F (1986) Botulinum A neurotoxin unlike tetanus toxin acts via a neuraminidase sensitive structure. Toxicon 24:1065–1074
Marxen P, Bigalke, H (1989) Tetanus toxin: inhibitory action in chromaffin cells is initiated by specified types of gangliosides and promoted in low ionic strength solution. Neurosci Lett 107:261–266
Marxen P, Fuhrmann U, Bigalke H (1989) Gangliosides mediate inhibitory effects of tetanus and botulinum A neurotoxins on exocytosis in chromaffin cells. Toxicon 27:849–859
Kamata Y, Yoshimoto M, Kozaki S (1997) Interaction between botulinum neurotoxin type A and ganglioside: ganglioside inactivates the neurotoxin and quenches its tryptophan fluorescence. Toxicon 35:1337–1340
Halpern JL, Loftus A (1993) Characterization of the receptor-binding domain of tetanus toxin. Nature 268:11188–11192
Shapiro RS, Specht CD, Collins BE, Woods AS, Cotter RJ, Schnaar RL (1997) Identification of a ganglioside recognition domain of tetanus toxin using a novel ganglioside photoaffinity ligand. J Biol Chem 272:30380–30386
Rummel A, Mahrhold S, Bigalke H, Binz T (2004) The HCC-domain of botulinum neurotoxins A and B exhibits a singular ganglioside binding site displaying serotype specific carbohydrate interaction. Mol Microbiol 51:631–644
Rummel A, Bade S, Alves J, Bigalke H, Binz T (2003) Two carbohydrate binding sites in the H(CC)-domain of tetanus neurotoxin are required for toxicity. J Mol Biol 326:835–847
Lazarovici P, Yavin E (1986) Affinity-purified tetanus neurotoxin interaction with synaptic membranes: properties of a protease-sensitive receptor component. Biochem Cell Biol 25:7047–7054
Pierce EJ, VDavison MD, Parton RG, Habig WJ, Cruitchley DR (1986) Characterization of tetanus toxin binding to rat brain membranes. Evidence for a high-affinity proteinase-sensitive receptor. Biochem J 236:845–852
Dong M, Yeh F, Tepp WH, Dean C, Johnson EA, Janz R, Chapman, ER (2006) SV2 is the protein receptor for botulinum neurotoxin A. Science 312:592–596
Dong M, Richards DA, Goodnough MC, Tepp WH, Johnson EA, Chapman ER (2003) Synaptotagmins I and II mediate entry of botulinum neurotoxin B into cells. J Cell Biol 162:1293–1303
Rummel A, Eichner T, Weil T, Karnath T, Gutcaits A, Mahrhold S, Sandhoff K, Proia RL, Acharya KR, Bigalke H, Binz T (2007) Identification of the protein receptor binding site of botulinum neurotoxins B and G proves the double-receptor concept. Proc Natl Acad Sci U S A 104:359–364
Dong M, Tepp WH, Liu H, Johnson EA, Chapman ER (2007) Mechanism of botulinum neurotoxin B and G entry into hippocampal neurons. J Cell Biol 179:1511–1522
Dong M, Liu H, Tepp WH, Johnson EA, Janz R, Chapman ER (2008) Glycosylated SV2A and SV2B mediate the entry of botulinum neurotoxin E into neurons. Mol Biol Cell 19:5226–5237
Eswaramoorthy S, Kumaran D, Keller J, Swaminathan S (2004) Role of metals in the biological activity of Clostridium botulinum neurotoxins. Biochemistry 43:2209–2216
Koriazova L, Montal M (2003) Translocation of botulinum neurotoxin light chain protease through the heavy chain channel. Nat Struct Biol 10:13–18
Fischer A, Montal M (2007) Crucial role of the disulfide bridge between botulinum neurotoxin light and heavy chains in protease translocation across membranes. J Biol Chem 282:29604–11.
Masuyer G, Thiyagarajan N, James PL, Marks PHH, Chaddock J, Acharya KR (2009) Crystal structure of a catalytically active, non-toxic endopeptidase derivative of Clostridium botulinum toxin A. Biochem Biophys Res Commun 381:50–53
Fischer A, Mushrush DJ, Lacy DB, Montal M (2008) Botulinum neurotoxin devoid of receptor binding domain translocates active protease. PLoS Pathog 4:e1000245
Montal M (2009) Translocation of botulinum neurotoxin light chain protease by the heavy chain protein-conducting channel. Toxicon 54:565–569
Galloux M, Vitrac H, Montagner C, Raffestin S, Popoff MR, Chenal A, Forge V, Gillet D (2008) Membrane Interaction of botulinum neurotoxin A translocation (T) domain. The belt region is a regulatory loop for membrane interaction. J Biol Chem 283:27668–27676
Agarwal R, Schmidt JJ, Stafford RG, Swaminathan S (2009) Mode of VAMP substrate recognition and inhibition of Clostridium botulinum neurotoxin F. Nat Struct Mol Biol 16:789–794
Brunger AT, Breidenbach MA, Jin R, Fischer A, Santos JS, Montal M (2007) Botulinum neurotoxin heavy chain belt as an intramolecular chaperone for the light chain. PLoS Pathog 3:1191–1194
Silvaggi NR, Wilson D, Tzipori S, Allen KN (2008) Catalytic features of the botulinum neurotoxin a light chain revealed by high resolution structure of an inhibitory peptide complex. Biochemistry 47:5736–5745
Kumaran D, Rawat R, Ahmed SA, Swaminathan S (2008) Substrate binding mode and its implication on drug design for botulinum neurotoxin A. PLoS Pathog 4:e1000165
Kumaran D, Rawat R, Ludivico ML, Ahmed SA, Swaminathan S (2008) Structure and substrate based inhibitor design for clostridium botulinum neurotoxin serotype A. J Biol Chem 283:18883–18891
Agarwal R, Binz T, Swaminathan S (2005) Analysis of active site residues of botulinum neurotoxin E by mutational, functional and structural studies: Glu335Gln is an apoenzyme. Biochemistry 44:8291–8302
Agarwal R, Binz T, Swaminathan S (2005) Structural analysis of botulinum neurotoxin serotype F light chain: implications on substrate binding and inhibitor design. Biochemistry 44:11758–11765
Binz T, Bade S, Rummel A, Kollewe A, Alves J (2002) Arg362 and Tyr365 of the botulinum neurotoxin type A light chain are involved in transition state stabilization. Biochemistry 41:1717–1723
Li L, Binz T, Niemann H, Singh BR (2000) Probing the mechanistic role of glutamate residues in the zinc-binding motif of type A botulinum neurotoxin light chain. Biochemistry 39:2399–2405
Li Y, Foran P, Fairweather NF, de Paiva A, Weller U, Dougan G, Dolly JO (1994) A single mutation in the recombinant light chain of tetanus toxin abolishes its proteolytic activity and removes the toxicity seen after reconstitution with native heavy chain. Biochemistry 33:7014–7020
Breidenbach MA, Brunger A (2004) Substrate recognition strategy for botulinum neurotoxin serotype A. Nature 432:925–929
Chen S, Barbieri JT (2006) Unique substrate recognition by botulinum neurotoxins serotypes A and E. J Biol Chem 281:10906–10911
Agarwal R, Swaminathan S (2008) SNAP-25 substrate peptide (residues 180–183) binds to but bypasses cleavage by catalytically active Clostridium botulinum neurotoxin E. J Biol Chem 283:25944–25951
Swaminathan S, Eswaramoorthy S, Kumaran D (2004) Structure and enzymatic activity of botulinum neurotoxins. Movement Disord 19(Suppl 8):S17–S22
Sikorra S, Henke T, Galli T, Binz T (2008) Substrate recognition mechanism of VAMP/synaptobrevin-cleaving clostridial neurotoxins. J Biol Chem 283:21145–21152
Schmidt JJ, Stafford RG (2005) Botulinum neurotoxin serotype F: identification of substrate recognition requirements and development of inhibitors with low nanomolar affinity. Biochemistry 44:4067–4073
Baldwin MR, Kim JJ, Barbieri JT (2007) Botulinum neurotoxin B-host receptor recognition: it takes two receptors to tango. Nat Struct Mol Biol 14:9–10
Wang J, Meng J, Lawrence GW, Zurawski TH, Sasse A, Bodeker MO, Gilmore MA, Fernandez-Salas E, Francis J, Steward LE, Aoki KR, Dolly JO (2008) Novel chimeras of botulinum neurotoxin/A and/E unveil contributions from the binding, translocation and protease domains to their functional characteristics. J Biol Chem 283:16993–17002
Marks JD (2004) Deciphering antibody properties that lead to potent botulinum neurotoxin neutralization. Mov Disord (Suppl 8):S101–S108
Lightstone FC, Prieto MC, Singh AK, Piqueras MC, Whittal RM, Knapp MS, Balhorn R, Roe DC (2000) Identification of novel small molecule ligands that bind to tetanus toxin. Chem Res Toxicol 13:356–362
Eswaramoorthy S, Kumaran D, Swaminathan S (2001) Crystallographic evidence for doxorubicin binding to the receptor-binding site in Clostridium botulinum neurotoxin B. Acta Crystallogr D Biol Crystallogr 57:1743–1746
Zou J, Miao WY, Ding FH, Meng JY, Ye HJ, Jia GR, He XY, Sun GZ, Li PZ (1985) The effect of toosendanin on monkey botulism. J Tradit Chin Med 5:29–30
Fischer A, Nakai Y, Eubanks LM, Clancy CM, Tepp WH, Pellett S, Dickerson TJ, Johnson EA, Janda KD, Montal M (2009) Bimodal modulation of the botulinum neurotoxin protein-conducting channel. Proc Natl Acad Sci U S A 106:1330–1335
Burnett JC, Henchal EA, Schmaljohn AL, Bavari S (2005) The evolving field of biodefence: therapeutic developments and diagnostics. Nat Rev Drug Discov 4:281–297
Burnett JC, Opsenica D, Sriraghavan K, Panchal RG, Ruthel G, Hermone AR, Nguyen TL, Kenny TA, Lane DJ, McGrath CF, Schmidt JJ, Vennerstrom JL, Gussio R, Solaja BA, Bavari S (2007) A refined pharmacophore identifies potent 4-amino-7-chloroquinoline-based inhibitors of the botulinum neurotoxin serotype A metalloprotease. J Med Chem 50:2127–2136
Burnett JC, Ruthel G, Stegmann CM, Panchal RG, Nguyen TL, Hermone AR, Stafford RG, Lane DJ, Kenny TA, McGrath CF, Wipf P, Stahl AM, Schmidt JJ, Gussio R, Brunger AT, Bavari S (2007) Inhibition of metalloprotease botulinum serotype A from a pseudo-peptide binding mode to a small molecule that is active in primary neurons. J Biol Chem 282, 5004–5014
Burnett JC, Schmidt JJ, McGrath CF, Nguyen TL, Hermone AR, Panchal RG, Vennerstrom JL, Kodukula K, Zaharevitz DW, Gussio R, Bavari S (2005) Conformational sampling of the botulinum neurotoxin serotype A light chain: implications for inhibitor binding. Bioorg Med Chem 13:333–341
Burnett JC, Schmidt JJ, Stafford RG, Panchal RG, Nguyen TL, Hermone AR, Vennerstrom JL, McGrath CF, Lane DJ, Sausville EA, Zaharevitz DW, Gussio R, Bavari S (2003) Novel small molecule inhibitors of botulinum neurotoxin A metalloprotease activity. Biochem Biophys Res Commun 310:84–93
Pang YP, Vummenthala A, Mishra RK, Park JG, Wang S, Davis J, Millard CB, Schmidt JJ (2010) Potent new small-molecule inhibitor of botulinum neurotoxin serotype A endopeptidase developed by synthesis-based computer-aided molecular design. PloS One 4:e7730
Tang J, Park JG, Millard CB, Schmidt JJ, Pang YP (2007) Computer-aided lead optimization: improved small-molecule inhibitor of the zinc endopeptidase of botulinum neurotoxin serotype A. PloS One 2:e761
Lai H, Feng M, Roxas-Duncan V, Dakshinamurthy S, Smith LA, Yang DC (2009) Quinolinol and peptide inhibitors of zinc protease in botulinum neurotoxin A: effects of zinc ion and peptides on inhibition. Arch Biochem Biophys 491:75–84
Roxas-Duncan V, Enyedy I, Montgomery VA, Eccard VS, Carrington MA, Lai H, Gul N, Yang DC, Smith LA (2009) Identification and biochemical characterization of small-molecule inhibitors of Clostridium botulinum neurotoxin serotype A. Antimicrob Agents Chemother 53:3478–3486
Capkova K, salzameda NT, Janda KD (2009) Investigations into small molecule non-peptidic inhibitors of the botulinum neurotoxins. Toxicon 54:575–582
Silvaggi NR, Boldt GE, Hixon MS, Kennedy JP, Tzipori S, Janda KD, Allen KN (2007) Structures of Clostridium botulinum neurotoxin serotype A light chain complexed with small-molecule inhibitors highlight active-site flexibility. Chem Biol 14:533–542
Zuniga JE, Hammill JT, Drory O, Nuss JE, Burnett JC, Gussio R, Wipf P, Bavari S, Brunger AT (2010) Iterative structure-based peptide-like inhibitor design against the botulinum neurotoxin serotype A. Plos One 5:e11378
Zuniga JE, Schmidt JJ, Fenn T, Burnett JC, Arac D, Gussio R, Stafford RG, Badie SS, Bavari S, Brunger AT (2008) A potent peptidomimetic inhibitor of botulinum neurotoxin serotype A has a very different conformation than SNAP-25 substrate. Structure (Camb) 16:1588–1597
Hale M, Oyler G, Swaminathan S, Ahmed SA (2010) Basic tetrapeptides as potent intracellular inhibitors of type A botulinum neurotoxin protease activity. J. Biol. Chem. 286:1802–1811
Schmidt JJ, Stafford RG (2002) A high-affinity competitive inhibitor of type A botulinum neurotoxin protease activity. FEBS Lett 532:423–426
Chen S, Barbieri JT (2009) Engineering botulinum neurotoxin to extend therapeutic intervention. Proc Natl Acad Sci 106:9180–9184
The author thanks his colleagues Drs. S. Eswaramoorthy, D. Kumaran, R. Agarwal, and G. Kumar and his collaborators for contributing to this research. Research was supported by an award from DTRA BO742081 under DOE prime contract No. DEAC02-98CH10886 with Brookhaven National Laboratory.
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Swaminathan, S. (2014). Neurotoxin Structure. In: Foster, K. (eds) Molecular Aspects of Botulinum Neurotoxin. Current Topics in Neurotoxicity, vol 4. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-9454-6_5
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