Crystal and Electron Microscopy Structures of Sticholysin II Actinoporin Reveal Insights into the Mechanism of Membrane Pore Formation

Mancheño, José M.; Martín‐Benito, Jaime; Martı́nez-Ripoll, M.; Gavilanes, José G.; Hermoso, J.A.

doi:10.1016/j.str.2003.09.019

Cited by 217 publications

(400 citation statements)

References 47 publications

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“…Considering previous experimental results on structure and conformation of actinoporins (Álvarez et al 1998, Miles et al 2008, Ulrih et al 2004, Mancheño et al 2001, 2003, Hinds et al 2002, Athanasiadis et al 2001, Poklar et al 1997, Malavasic et al 1996, six systems were submitted to MD simulations: 1) EqtxII under pH = 7.0; 2) EqtxII under pH = 3.0; 3) StnII under pH = 7.0; 4) StnII under pH = 3.0; 5) StnII under pH = 7.0 starting from the last frame of the simulation at pH = 3.0; 6) StnII in 10.0 M urea solution. While Systems 1-4 and 6 employed, as their starting geometries, the respective StnII and EqtxII crystallographic structures, System 5 was simulated using the last frame from System 4 as its starting geometry.…”

Section: Simulation Systemsmentioning

confidence: 94%

“…These toxins have a remarkable sequence similarity (higher than 90%) and present a globular shape with a β-sandwich fold, flanked by N-and C-terminal α-helices (Mancheño et al 2003, Hinds et al 2002, Athanasiadis et al 2001, Huerta et al 2001. Their structure and conformation have been studied by different approaches (Norton 2009), including circular dichroism (CD) , Poklar et al 1997, fluorescence measurements (Álvarez et al , Malavasic et al 1996, X-ray crystallography (Mancheño et al 2003, Athanasiadis et al 2001) and NMR spectroscopy (Hinds et al 2002) (Figure 1).…”

Section: Introductionmentioning

confidence: 99%

“…Their structure and conformation have been studied by different approaches (Norton 2009), including circular dichroism (CD) , Poklar et al 1997, fluorescence measurements (Álvarez et al , Malavasic et al 1996, X-ray crystallography (Mancheño et al 2003, Athanasiadis et al 2001) and NMR spectroscopy (Hinds et al 2002) (Figure 1). …”

Section: Introductionmentioning

confidence: 99%

“…They are: the phosphocholine binding site (Ser52,Val85,Ser103,Pro105,Tyr111,Tyr131,Tyr135,Tyr136 in StnII and Ser54,Val87,Ser105,Pro107,Tyr113,Tyr133,Tyr137,Tyr138 in EqtxII), the tryptophan-rich region (Trp110,Trp114,Trp115 in StnII and Trp112,Trp116,Trp117 in EqtxII), and the N-termini (Ala1-Ser30 in StnII and Ser1-Ile33 in EqtxII) (Mancheño et al 2003, Hong et al 2002, Anderluh et al 1997.…”

Section: Introductionmentioning

confidence: 99%

“…It is proposed that the phosphocholine binding site is responsible for the recognition of its molecular target and interaction between the Trp-rich region and the lipid bilayers (Mancheño et al 2003, Hong et al 2002). Such event is followed by an N-terminal displacement into the lipid bilayer, thought to be mandatory for the functional pore formation (Casallanovo et al 2006, Kristan et al 2004).…”

Section: Introductionmentioning

confidence: 99%

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Differential Effect of Solution Conditions on the Conformation of the Actinoporins Sticholysin II and Equinatoxin II

Fauth

Cilli

Ligabue-Braun

et al. 2014

An. Acad. Bras. Ciênc.

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Actinoporins are a family of pore-forming proteins with hemolytic activity. The structural basis for such activity appears to depend on their correct folding. Such folding encompasses a phosphocholine binding site, a tryptophan-rich region and the activity-related N-terminus segment. Additionally, different solution conditions are known to be able to influence the pore formation by actinoporins, as for Sticholysin II (StnII) and Equinatoxin II (EqtxII). In this context, the current work intends to characterize the influence of distinct solution conditions in the conformational behavior of these proteins through molecular dynamics (MD) simulations. The obtained data offer structural insights into actinoporins dynamics in solution, characterizing its conformational behavior at the atomic level, in accordance with previous experimental data on StnII and EqtxII hemolytic activities.

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Section: Simulation Systemsmentioning

confidence: 94%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Differential Effect of Solution Conditions on the Conformation of the Actinoporins Sticholysin II and Equinatoxin II

Fauth

Cilli

Ligabue-Braun

et al. 2014

An. Acad. Bras. Ciênc.

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Pore‐forming Toxins

Serra

Martínez

2011

Encyclopedia of Life Sciences

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Pore‐forming toxins (PFT) are proteins able to produce well‐structured holes in target cell membrane. They have a very broad taxonomic distribution being produced from bacteria to mammals. Depending on the secondary structure of the membrane‐spanning region, these proteins are categorised into two classes: α‐PFT and β‐PFT. The pore structure of representative members of each class will be described. These proteins can be also classified according to their pore structure: barrel‐stave and toroidal protein–lipid pore. In the barrel‐stave pore the protein molecules provide a continuous interface between the core of the bilayer and the channel lumen, whereas in the toroidal protein–lipid pore both polypeptide chains and polar phospholipid headgroups are involved in the building of pore walls. The stoichiometry and the pore diameter depend on the protein, thus the channels can allow leakage of ions, adenosine triphosphate (ATP), proteins and even bacteria. Attacked cells trigger different responses, some promoting recovery of membrane integrity, others transition to a low‐energy‐consumption state, in addition to inflammatory responses and changes in gene transcription. Key Concepts: Pore‐forming toxins are proteins produced from bacteria to mammals. Pore‐forming toxins are classified as α‐PFT and β‐PFT, according to the structure adopted by the transmembrane region. Pores are nanometre funnel‐like holes that permit the passage of water, ions and small molecules through cell membranes. PFT form two types of pores, the barrel‐stave and the toroidal protein–lipid pore. Cells react to pore formation, repair the damage and survive.

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Model peptides mimic the structure and function of the N‐terminus of the pore‐forming toxin sticholysin II

Casallanovo¹,

Oliveira²,

Souza³

et al. 2005

Biopolymers

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To investigate the role of the N-terminal region in the lytic mechanism of the pore-forming toxin sticholysin II (St II), we studied the conformational and functional properties of peptides encompassing the first 30 residues of the protein. Peptides containing residues 1-30 (P1-30) and 11-30 (P11-30) were synthesized and their conformational properties were examined in aqueous solution as a function of peptide concentration, pH, ionic strength, and addition of the secondary structure-inducing solvent trifluoroethanol (TFE). CD spectra showed that increasing concentration, pH, and ionic strength led to aggregation of P1-30; as a consequence, the peptide acquired beta-sheet conformation. In contrast, P11-30 exhibited practically no conformational changes under the same conditions, remaining essentially structureless. Moreover, this peptide did not undergo aggregation. These differences clearly point to the modulating effect of the first 10 hydrophobic residues on the peptides aggregation and conformational properties. In TFE both the first ten hydrophobic peptides acquired alpha-helical conformation, albeit to a different extent, P11-30 displayed lower alpha-helical content. P1-30 presented a larger fraction of residues in alpha-helical conformation in TFE than that found in St II's crystal structure for that portion of the protein. Since TFE mimics the membrane environment, such increase in helical content could also occur upon toxin binding to membranes and represent a step in the mechanism of pore formation. The peptides conformational properties correlated well with their functional behavior. Thus, P1-30 exhibited much higher hemolytic activity than P11-30. In addition, P11-30 was able to block the toxin's hemolytic activity. The size of pores formed in red blood cells by P1-30 was estimated by measuring the permeability to PEGs of different molecular mass. The pore radius (0.95 +/- 0.01 nm) was very similar to that of the pore formed by the toxin. The results demonstrate that the synthetic peptide P1-30 is a good model of St II conformation and function and emphasize the contribution of the toxin's N-terminal region, and, in particular, the hydrophobic residues 1-10 to pore formation.

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Crystal and Electron Microscopy Structures of Sticholysin II Actinoporin Reveal Insights into the Mechanism of Membrane Pore Formation

Cited by 217 publications

References 47 publications

Differential Effect of Solution Conditions on the Conformation of the Actinoporins Sticholysin II and Equinatoxin II

Differential Effect of Solution Conditions on the Conformation of the Actinoporins Sticholysin II and Equinatoxin II

Pore‐forming Toxins

Model peptides mimic the structure and function of the N‐terminus of the pore‐forming toxin sticholysin II

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