Pore-forming toxins (PFT) are water-soluble proteins that possess the remarkable ability to self-assemble on the membrane of target cells, where they form pores causing cell damage. Here, we elucidate the mechanism of action of the haemolytic protein fragaceatoxin C (FraC), a α-barrel PFT, by determining the crystal structures of FraC at four different stages of the lytic mechanism, namely the water-soluble state, the monomeric lipid-bound form, an assembly intermediate and the fully assembled transmembrane pore. The structure of the transmembrane pore exhibits a unique architecture composed of both protein and lipids, with some of the lipids lining the pore wall, acting as assembly cofactors. The pore also exhibits lateral fenestrations that expose the hydrophobic core of the membrane to the aqueous environment. The incorporation of lipids from the target membrane within the structure of the pore provides a membrane-specific trigger for the activation of a haemolytic toxin.
Plinabulin (11, NPI-2358) is a potent microtubule-targeting agent derived from the natural diketopiperazine "phenylahistin" (1) with a colchicine-like tubulin depolymerization activity. Compound 11 was recently developed as VDA and is now under phase II clinical trials as an anticancer drug. To develop more potent antimicrotubule and cytotoxic derivatives based on the didehydro-DKP skeleton, we performed further modification on the tert-butyl or phenyl groups of 11, and evaluated their cytotoxic and tubulin-binding activities. In the SAR study, we developed more potent derivatives 33 with 2,5-difluorophenyl and 50 with a benzophenone in place of the phenyl group. The anti-HuVEC activity of 33 and 50 exhibited a lowest effective concentration of 2 and 1 nM for microtubule depolymerization, respectively. The values of 33 and 50 were 5 and 10 times more potent than that of CA-4, respectively. These derivatives could be a valuable second-generation derivative with both vascular disrupting and cytotoxic activities.
Previous studies have demonstrated improvement of regional wall motion and global left ventricular function after successful recanalization of chronic total occlusion in coronary artery. However, the difference of benefits of recanalization between infarct site and noninfarct site is unknown. This study assessed the changes in left ventricular ejection fraction, regional wall motion after successful angioplasty of chronic total occlusions with or without previous myocardial infarction. This study also evaluated the factors that influenced the outcome of left ventricular function. We retrospectively studied 75 patients with a successfully recanalized chronic total occlusion in native coronary artery. Left ventriculograms were obtained at baseline and after 6 months. Global and regional left ventricular function were determined. The patients were divided into two groups. Group 1 comprised patients without previous myocardial infarction in the territories of total occlusion vessel that was recanalized. Group 2 comprised patients with previous myocardial infarction in the territories of total occlusion vessel that was recanalized. Left ventricular ejection fraction increased from 53.2% +/- 16.3% at baseline to 57.3% +/- 20.1% at 6-month follow-up in the whole group (P = 0.001). In group 1 patients, the evolution of left ventricular (LV) ejection fraction increased from 59.5% +/- 13.7% to 67.3% +/- 14.6% (P < 0.001). In group 2 patients, the evolution of LV ejection fraction increased, but not significantly, from 48.9% +/- 16.2% to 50.5% +/- 16.9% (P = NS). The evolution of LV ejection fraction increased from 47.6% +/- 17.4% to 50.8% +/- 17.5% (P < 0.05) in the subgroup of recanalization in infarct-related vessel that had rich collateral circulation and had long-term patency. The regional wall motion all significantly improved in group 1 patients (P < 0.05). The regional wall motion did not change in group 2 patients (P = NS). The influence of recanalization of chronic coronary occlusions on the improvement of left ventricular global function was different between myocardial infarction and nonmyocardial infarction patients. The left ventricular function did not improve in myocardial infarction patient. Regional wall motion improved in patients without previous myocardial infarction. For reliable improvement of left ventricular function after recanalization of chronic total occlusions, evidence (not only by symptom or treadmill test) of viable myocardium in recanalized vessel is important. It is also important to keep patency of infarct-related vessel that has good collateral circulation for improving the left ventricular function.
This paper describes the analysis of pore formation and detection of a single protein molecule using a large nanopore among five different pore-forming proteins. We demonstrate that the identification of appropriate pores for nanopore sensing can be achieved by classifying the channel current signals and performing noise analysis. Through these analyses, we selected a perforin nanopore from the membrane attack complex/perforin superfamily and attempted to use it to detect the granzyme B protein, a serine protease. As a result, we found that granzyme B might pass through the perforin nanopore if it adopts an unfolded structure. Our proposed analytical approach should be useful for exploring several types of nanopore as large biological nanopores other than α-hemolysin.
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