The interaction of baccatin III with the taxol binding site of microtubules determined by a homogeneous assay with fluorescent taxoid.
AuthorsAndreu JM, Barasoain I
JournalBiochemistry
PubMed ID11580273
'The ubiquitous Taxol binding site of microtubules also binds newly discovered ligands. We have designed a homogeneous assay for the high throughput detection of Taxol biomimetics, based on the displacement of 7-O-[N-(2,7-difluoro-4''-fluoresceincarbonyl)-L-alanyl]Taxol from its binding site in diluted solutions of preserved microtubules. The state of this reference ligand is measured ... More
An incremental approach to automated protein localisation.
AuthorsTscherepanow M, Jensen N, Kummert F,
JournalBMC Bioinformatics
PubMed ID18937856
'BACKGROUND: The subcellular localisation of proteins in intact living cells is an important means for gaining information about protein functions. Even dynamic processes can be captured, which can barely be predicted based on amino acid sequences. Besides increasing our knowledge about intracellular processes, this information facilitates the development of innovative ... More
Molecular recognition of taxol by microtubules. Kinetics and thermodynamics of binding of fluorescent taxol derivatives to an exposed site.
AuthorsDíaz JF, Strobe R, Engelborghs Y, Souto AA, Andreu JM
JournalJ Biol Chem
PubMed ID10818101
'We have determined the kinetic scheme and the reaction rates of binding to microtubules of two fluorescent taxoids, 7-O-[N-(4''-fluoresceincarbonyl)-l-alanyl]Taxol (Flutax-1) and 7-O-[N-(2,7-difluoro-4''-fluoresceincarbonyl)-l-alanyl]Taxol (Flutax-2). Flutax-1 and Flutax-2 bind to microtubules with high affinity (K(a) approximately 10(7) m(-1), 37 degrees C). The binding mechanism consists of a fast bimolecular reaction followed by ... More
Fluorescent taxoid probes for microtubule research.
AuthorsBarasoain I, Díaz JF, Andreu JM,
JournalMethods Cell Biol
PubMed ID20466144
'The use of the antitumor drug taxol as an experimental microtubule-stabilizing agent is widespread. Fluorescent taxol conjugates, although less employed, are very useful tools for several purposes in microtubule research. These include easily visualizing microtubule cytoskeletons in a variety of cells as well as in vitro assembled microtubules, studying the ... More
Perspective of a new diagnostic for human trichomonosis.
AuthorsLecke SB, Tasca T, Souto AA, De Carli GA
JournalMem Inst Oswaldo Cruz
PubMed ID12764446
Several diagnostic techniques have been employed for the detection of Trichomonas vaginalis. Microtubules constitute the cytoskeleton in eukaryotic cells and are sensitive to antimitotic drugs, such as Taxol (paclitaxel). We used FLUTAX a fluorescent taxoid - to analyze the microtubule distribution in living trophozoites of T. vaginalis in urine and ... More
In vitro formation of the endoplasmic reticulum occurs independently of microtubules by a controlled fusion reaction.
AuthorsDreier L, Rapoport TA
JournalJ Cell Biol
PubMed ID10704440
We have established an in vitro system for the formation of the endoplasmic reticulum (ER). Starting from small membrane vesicles prepared from Xenopus laevis eggs, an elaborate network of membrane tubules is formed in the presence of cytosol. In the absence of cytosol, the vesicles only fuse to form large ... More
Biodegradable polymersomes as carriers and release systems for paclitaxel using Oregon Green® 488 labeled paclitaxel as a model compound.
AuthorsLee JS, Feijen J,
JournalJ Control Release
PubMed ID22063005
Oregon Green® 488 labeled paclitaxel (Flutax) loaded biodegradable polymersomes (Flutax-Ps) based on methoxy poly(ethylene glycol)-b-poly(d,l-lactide) (mPEG-PDLLA), methoxy poly(ethylene glycol)-b-poly(e-caprolactone) (mPEG-PCL) or a mixture of the block copolymers (50:50, w/w) were prepared (abbreviated as Flutax-Ps (L), Flutax-Ps (C) and Flutax-Ps (LC), respectively). For the formation of the Ps, the corresponding block ... More
The plant formin AtFH4 interacts with both actin and microtubules, and contains a newly identified microtubule-binding domain.
AuthorsDeeks MJ, Fendrych M, Smertenko A, Bell KS, Oparka K, Cvrcková F, Zársky V, Hussey PJ,
JournalJ Cell Sci
PubMed ID20332108
The dynamic behaviour of the actin cytoskeleton in plants relies on the coordinated action of several classes of actin-binding proteins (ABPs). These ABPs include the plant-specific subfamilies of actin-nucleating formin proteins. The model plant species Arabidopsis thaliana has over 20 formin proteins, all of which contain plant-specific regions in place ... More
Myosin XI-dependent formation of tubular structures from endoplasmic reticulum isolated from tobacco cultured BY-2 cells.
AuthorsYokota E, Ueda H, Hashimoto K, Orii H, Shimada T, Hara-Nishimura I, Shimmen T,
JournalPlant Physiol
PubMed ID21427277
The reticular network of the endoplasmic reticulum (ER) consists of tubular and lamellar elements and is arranged in the cortical region of plant cells. This network constantly shows shape change and remodeling motion. Tubular ER structures were formed when GTP was added to the ER vesicles isolated from tobacco (Nicotiana ... More
Application of ultrasound to selectively localize nanodroplets for targeted imaging and therapy.
AuthorsDayton PA, Zhao S, Bloch SH, Schumann P, Penrose K, Matsunaga TO, Zutshi R, Doinikov A, Ferrara KW,
JournalMol Imaging
PubMed ID16954031
Lipid-coated perfluorocarbon nanodroplets are submicrometer-diameter liquid-filled droplets with proposed applications in molecularly targeted therapeutics and ultrasound (US) imaging. Ultrasonic molecular imaging is unique in that the optimal application of these agents depends not only on the surface chemistry, but also on the applied US field, which can increase receptor-ligand binding ... More
Docking and rolling, a model of how the mitotic motor Eg5 works.
AuthorsRosenfeld SS, Xing J, Jefferson GM, King PH
JournalJ Biol Chem
PubMed ID16115880
Whereas kinesin I is designed to transport cargoes long distances in isolation, a closely related kinesin motor, Eg5, is designed to generate a sustained opposing force necessary for proper mitotic spindle formation. Do the very different roles for these evolutionarily related motors translate into differences in how they generate movement? ... More
Correct diffusion coefficients of proteins in fluorescence correlation spectroscopy. Application to tubulin oligomers induced by Mg2+ and Paclitaxel.
AuthorsKrouglova T, Vercammen J, Engelborghs Y
JournalBiophys J
PubMed ID15454458
In view of recent warnings for artifacts in fluorescence correlation spectroscopy, the diffusion coefficient of a series of labeled proteins in a wide range of molecular mass (43-670 kD) was determined and shown to be correct with respect to published values and the theory. Fluorescence correlation spectroscopy was then applied ... More
Fast kinetics of Taxol binding to microtubules. Effects of solution variables and microtubule-associated proteins.
AuthorsDíaz JF, Barasoain I, Andreu JM
JournalJ Biol Chem
PubMed ID12496245
The kinetics of Taxol association to and dissociation from stabilized microtubules has been measured by competition with the reference fluorescent derivative Flutax-1 (Diaz, J. F., Strobe, R., Engelborghs, Y., Souto, A. A., and Andreu, J. M. (2000) J. Biol. Chem. 275, 26265-26276). The association rate constant at 37 degrees C ... More
Microtubule bundling and nested buckling drive stripe formation in polymerizing tubulin solutions.
AuthorsLiu Y, Guo Y, Valles JM, Tang JX
JournalProc Natl Acad Sci U S A
PubMed ID16818889
Various mechanisms govern pattern formation in chemical and biological reaction systems, giving rise to structures with distinct morphologies and physical properties. The self-organization of polymerizing microtubules (MTs) is of particular interest because of its implications for biological function. We report a study of the microscopic structure and properties of the ... More
Paclitaxel-resistant human ovarian cancer cells undergo c-Jun NH2-terminal kinase-mediated apoptosis in response to noscapine.
AuthorsZhou J, Gupta K, Yao J, Ye K, Panda D, Giannakakou P, Joshi HC
JournalJ Biol Chem
PubMed ID12183452
We have previously discovered the opium alkaloid noscapine as a microtubule interacting agent that binds to tubulin, alters the dynamics of microtubule assembly, and arrests mammalian cells at mitosis (Ye, K., Ke, Y., Keshava, N., Shanks, J., Kapp, J. A., Tekmal, R. R., Petros, J., and Joshi, H. C. (1998) ... More
Location and properties of the taxol binding center in microtubules: a picosecond laser study with fluorescent taxoids.
AuthorsLillo MP, Cañadas O, Dale RE, Acuña AU
JournalBiochemistry
PubMed ID12369834
The interaction of two bioactive, fluorescent analogues of the anticancer drug Taxol, Flutax1 [7-O-[N-(fluorescein-4'-carbonyl)-L-alanyl]taxol] and Flutax2 [7-O-[N-(2,7-difluorofluorescein-4'-carbonyl)-L-alanyl]taxol], with microtubules in solution has been studied with picosecond laser methods. As shown here, although a mixture of the fluorescein mono- and dianion species of Flutax1 is present in solution, the bound taxoid ... More
Dissecting cellular processes using small molecules: identification of colchicine-like, taxol-like and other small molecules that perturb mitosis.
BACKGROUND: Understanding the molecular mechanisms of complex cellular processes requires unbiased means to identify and to alter conditionally gene products that function in a pathway of interest. Although random mutagenesis and screening (forward genetics) provide a useful means to this end, the complexity of the genome, long generation time and ... More
Centrosome and spindle pole microtubules are main targets of a fluorescent taxoid inducing cell death.
AuthorsAbal M, Souto AA, Amat-Guerri F, Acuña AU, Andreu JM, Barasoain I
JournalCell Motil Cytoskeleton
PubMed ID11309836
Microtubules offer a very large local concentration of binding sites for cytotoxic taxoids or for hypothetical endogenous regulators. Several compounds from diverse sources stabilize microtubules and arrest cell division similarly to the antitumour drug Taxol. We have investigated the subcellular location of the Taxol binding sites, employing a fluorescent taxoid ... More
Engineered human blood-brain barrier microfluidic model for vascular permeability analyses.
Authors
JournalNat Protoc
PubMed ID34997242
A nanoparticle formula for delivering siRNA or miRNAs to tumor cells in cell culture and in vivo.
Authors
JournalNat Protoc
PubMed ID25033207
Delivery of membrane impermeable cargo into CHO cells by peptide nanoparticles targeted by a protein corona.
Authors
JournalBiomaterials
PubMed ID22226586
Spatial confinement of active microtubule networks induces large-scale rotational cytoplasmic flow.
Authors
JournalProc Natl Acad Sci U S A
PubMed ID28265076
Single-molecule-based super-resolution images in the presence of multiple fluorophores.
Authors
JournalNano Lett
PubMed ID22003850
Paclitaxel-exposed ovarian cancer cells induce cancer‑specific CD4+ T cells after doxorubicin exposure through regulation of MyD88 expression.
Authors
JournalInt J Oncol
PubMed ID24573741
Multifunctional Polymeric Micelles Co-loaded with Anti-Survivin siRNA and Paclitaxel Overcome Drug Resistance in an Animal Model of Ovarian Cancer.
Authors
JournalMol Cancer Ther
PubMed ID25657335
Direct assessment of P-glycoprotein efflux to determine tumor response to chemotherapy.
Authors
JournalBiochem Pharmacol
PubMed ID20298675
Application of Transmural Flow Across In Vitro Microvasculature Enables Direct Sampling of Interstitial Therapeutic Molecule Distribution.
Authors
JournalSmall
PubMed ID31497931
Macropinocytosis of Nab-paclitaxel Drives Macrophage Activation in Pancreatic Cancer.
Authors
JournalCancer Immunol Res
PubMed ID28108630
Protein and chemotherapy profiling of extracellular vesicles harvested from therapeutic induced senescent triple negative breast cancer cells.
AuthorsKavanagh EL, Lindsay S, Halasz M, Gubbins LC, Weiner-Gorzel K, Guang MHZ, McGoldrick A, Collins E, Henry M, Blanco-Fernández A, O Gorman P, Fitzpatrick P, Higgins MJ, Dowling P, McCann A
JournalOncogenesis
PubMed ID28991260
Triple negative breast cancer (TNBC) is an aggressive subtype with relatively poor clinical outcomes and limited treatment options. Chemotherapy, while killing cancer cells, can result in the generation of highly chemoresistant therapeutic induced senescent (TIS) cells that potentially form stem cell niches resulting in metastases. Intriguingly, senescent cells release significantly ... More