Abstract：
:In the past few decades, marine natural products bioprospecting has yielded a considerable number of drug candidates. Two marine natural products have recently been admitted as new drugs: Prialt (also known as ziconotide) as a potent analgesic for severe chronic pain and Yondelis (known also as trabectedin or E-743) as antitumor agent for the treatment of advanced soft tissue sarcoma. In this protocol, methods for bioactivity-guided isolation, purification and identification of secondary metabolites from marine invertebrates such as sponges, tunicates, soft corals and crinoids are discussed. To achieve this goal, solvent extraction of usually freeze-dried sample of marine organisms is performed. Next, the extract obtained is fractionated by liquid-liquid partitioning followed by various chromatographic separation techniques including thin layer chromatography, vacuum liquid chromatography, column chromatography (CC) and preparative high-performance reversed-phase liquid chromatography. Isolation of bioactive secondary metabolites is usually monitored by bioactivity assays, e.g., antioxidant (2,2-diphenyl-1-picryl hydrazyl) and cytotoxicity (microculture tetrazolium) activities that ultimately yield the active principles. Special care should be taken when performing isolation procedures adapted to the physical and chemical characteristics of the compounds isolated, particularly their lipo- or hydrophilic characters. Examples of isolation of compounds of different polarities from extracts of various marine invertebrates will be presented in this protocol. Structure elucidation is achieved using recent spectroscopic techniques, especially 2D NMR and mass spectrometry analysis.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Ebada SS,Edrada RA,Lin W,Proksch Pdoi
10.1038/nprot.2008.182subject
Has Abstractpub_date
2008-01-01 00:00:00pages
1820-31issue
12eissn
1754-2189issn
1750-2799pii
nprot.2008.182journal_volume
3pub_type
杂志文章相关文献
Nature Protocols文献大全abstract：:Multipotent neural crest stem cells (NCSCs) have the potential to generate a wide range of cell types including melanocytes; peripheral neurons; and smooth muscle, bone, cartilage and fat cells. This protocol describes in detail how to perform a highly efficient, lineage-specific differentiation of human pluripotent c...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.156
更新日期：2013-01-01 00:00:00
abstract：:Human pluripotent stem cells (hPSCs) provide a valuable model for the study of human development and a means to generate a scalable source of cells for therapeutic applications. This protocol specifies cell fate efficiently into cardiac and endothelial lineages from hPSCs. The protocol takes 2 weeks to complete and re...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2016.153
更新日期：2017-01-01 00:00:00
abstract：:This protocol describes a general method for the preparation of RNAs in which the reactivity or hydrogen-bonding properties of the molecule are modified in a photoreversible fashion by use of a caging strategy. A single caged adenosine, modified at the 2' position as a nitro-benzyl ether, can be incorporated into shor...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.154
更新日期：2007-01-01 00:00:00
abstract：:Embryonic stem cells (ESCs) constitute a tool of great potential in neurobiology, enabling the directed differentiation of specific neural cell types. We have shown recently that neurons of the cerebral cortex can be generated from mouse ESCs cultured in a chemically defined medium that contains no morphogen, but in t...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2009.157
更新日期：2009-01-01 00:00:00
abstract：:Here we describe a protocol to dissect the peroxisomal matrix protein import pathway using a cell-free in vitro system. The system relies on a postnuclear supernatant (PNS), which is prepared from rat/mouse liver, to act as a source of peroxisomes and cytosolic components. A typical in vitro assay comprises the follow...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2016.147
更新日期：2016-12-01 00:00:00
abstract：:This protocol describes the synthesis of oligo(poly(ethylene glycol) fumarate) (OPF; 1-35 kDa; a polymer useful for tissue engineering applications) by a one-pot reaction of poly(ethylene glycol) (PEG) and fumaryl chloride. The procedure involves three parts: dichloromethane and PEG are first dried; the reaction step ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.055
更新日期：2012-05-31 00:00:00
abstract：:Ex vivo perfusion systems offer a reliable, reproducible method for studying acute physiological responses of an organ to various environmental manipulations. Unlike in vitro culture systems, the cellular organization, compartmentalization and three-dimensional structure of ex vivo-perfused organs are maintained. Thes...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.144
更新日期：2013-01-01 00:00:00
abstract：:The unprecedented increase in the throughput of DNA sequencing driven by next-generation technologies now allows efficient analysis of the complete protein-coding regions of genomes (exomes) for multiple samples in a single sequencing run. However, sample preparation and targeted enrichment of multiple samples has bec...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2011.396
更新日期：2011-11-03 00:00:00
abstract：:Iterative saturation mutagenesis (ISM) is a new and efficient method for the directed evolution of functional enzymes. It reduces the necessary molecular biological work and the screening effort drastically. It is based on a Cartesian view of the protein structure, performing iterative cycles of saturation mutagenesis...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.72
更新日期：2007-01-01 00:00:00
abstract：:Small-angle X-ray scattering (SAXS) and small-angle neutron scattering (SANS) are techniques used to extract structural parameters and determine the overall structures and shapes of biological macromolecules, complexes and assemblies in solution. The scattering intensities measured from a sample contain contributions ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2016.113
更新日期：2016-11-01 00:00:00
abstract：:The transmembrane (TM) anchors of cell surface proteins have been one of the 'blind spots' in structural biology because they are generally very hydrophobic, sometimes dynamic, and thus difficult targets for structural characterization. A plethora of examples show these membrane anchors are not merely anchors but can ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-019-0188-9
更新日期：2019-08-01 00:00:00
abstract：:Several cell death assays have been developed based on a single biochemical parameter such as caspase activation or plasma membrane permeabilization. Our fluorescent apoptosis/necrosis (FAN) assay directly measures cell death and distinguishes between caspase-dependent apoptosis and caspase-independent necrosis of cel...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2016.085
更新日期：2016-08-01 00:00:00
abstract：:The increasing multi-resistance of pathogenic bacteria requires the development of novel classes of antibiotics. Antimicrobial host defense peptides represent one promising class. Here we describe a protocol for screening large numbers of peptides against any microbe of interest. Peptides synthesized on a cellulose su...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.203
更新日期：2007-01-01 00:00:00
abstract：:We describe the use of a microfabricated cell culture substrate, consisting of a uniform array of closely spaced, vertical, elastomeric microposts, to study the effects of substrate rigidity on cell function. Elastomeric micropost substrates are micromolded from silicon masters comprised of microposts of different hei...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2010.189
更新日期：2011-02-01 00:00:00
abstract：:Raman spectroscopy can be used to measure the chemical composition of a sample, which can in turn be used to extract biological information. Many materials have characteristic Raman spectra, which means that Raman spectroscopy has proven to be an effective analytical approach in geology, semiconductor, materials and p...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2016.036
更新日期：2016-04-01 00:00:00
abstract：:Hydroxyl radical footprinting has been widely used for studying the structure of DNA and DNA-protein complexes. The high reactivity and lack of base specificity of the hydroxyl radical makes it an excellent probe for high-resolution footprinting of DNA-protein complexes; this technique can provide structural detail th...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2008.72
更新日期：2008-01-01 00:00:00
abstract：:Biominerals serve as critical structures of living systems and play important roles in biochemical processes. Understanding their crystallization mechanisms is therefore central to many areas of biology, biogeoscience, and biochemistry. Some biominerals, such as bone and dentin, are hierarchical nanocomposite structur...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-018-0027-4
更新日期：2018-09-01 00:00:00
abstract：:Determining enantiomeric excess (e.e.) in chiral compounds is key to development of chiral catalyst auxiliaries and chiral drugs. Here we describe a sensitive and robust fluorescence-based assay for determining e.e. in mixtures of enantiomers of 1,2- and 1,3-diols, chiral amines, amino alcohols, and amino-acid esters....
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-020-0329-1
更新日期：2020-07-01 00:00:00
abstract：:This protocol describes a culture system in which inner-ear sensory tissue is produced from mouse embryonic stem (ES) cells under chemically defined conditions. This model is amenable to basic and translational investigations into inner ear biology and regeneration. In this protocol, mouse ES cells are aggregated in 9...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2014.100
更新日期：2014-01-01 00:00:00
abstract：:This protocol details a subset of assays developed within the touchscreen platform to measure various aspects of executive function in rodents. Three main procedures are included: extinction, measuring the rate and extent of curtailing a response that was previously, but is no longer, associated with reward; reversal ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2013.123
更新日期：2013-10-01 00:00:00
abstract：:Here, we describe a simple micromolding method to construct three-dimensional arrays of organotypic epithelial tissue structures that approximate in vivo histology. An elastomeric stamp containing an array of posts of defined geometry and spacing is used to mold microscale cavities into the surface of type I collagen ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2008.35
更新日期：2008-01-01 00:00:00
abstract：:Here, we describe a robust protocol for the reverse transfection of cells on small interfering (siRNA) arrays, which, in combination with multi-channel immunofluorescence or time-lapse microscopy, is suitable for genome-wide RNA interference (RNAi) screens in intact human cells. The automatic production of 48 'transfe...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.483
更新日期：2007-01-01 00:00:00
abstract：:Glycoproteins are involved in diverse biological processes ranging from extracellular contact and recognition to intracellular signaling. Crystal structures of glycoproteins would yield tremendous insight into these processes. But glycoprotein structural analysis has been hindered by difficulties in expressing milligr...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2009.29
更新日期：2009-01-01 00:00:00
abstract：:We describe the implementation and use of an adaptive imaging framework for optimizing spatial resolution and signal strength in a light-sheet microscope. The framework, termed AutoPilot, comprises hardware and software modules for automatically measuring and compensating for mismatches between light-sheet and detecti...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-018-0043-4
更新日期：2018-11-01 00:00:00
abstract：:This protocol presents the peptide incorporation of environment-sensitive fluorophores derived from the dimethylaminophthalimide family. The procedure utilizes anhydride precursors of 4-dimethylaminophthalimide (4-DMAP) or 6-dimethylaminonaphthalimide (6-DMN), whose syntheses are described in a related protocol from t...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.442
更新日期：2007-01-01 00:00:00
abstract：:The adeno-associated virus (AAV) is one of the most promising viral vectors for human gene therapy. As with any potential therapeutic system, a thorough understanding of it at the in vitro and in vivo levels is required. Over the years, numerous methods have been developed to better characterize AAV vectors. These met...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.207
更新日期：2006-01-01 00:00:00
abstract：:The plant cell wall is a chemically complex structure composed mostly of polysaccharides. Detailed analyses of these cell wall polysaccharides are essential for our understanding of plant development and for our use of plant biomass (largely wall material) in the food, agriculture, fabric, timber, biofuel and biocompo...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.081
更新日期：2012-09-01 00:00:00
abstract：:The mouse embryo hindbrain is a robust and adaptable model for studying sprouting angiogenesis. It permits the spatiotemporal analysis of organ vascularization in normal mice and in mouse strains with genetic mutations that result in late embryonic or perinatal lethality. Unlike postnatal models such as retinal angiog...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2013.015
更新日期：2013-02-01 00:00:00
abstract：:Cognitive computing is revolutionizing the way big data are processed and integrated, with artificial intelligence (AI) natural language processing (NLP) platforms helping researchers to efficiently search and digest the vast scientific literature. Most available platforms have been developed for biomedical researcher...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-020-00455-4
更新日期：2021-01-22 00:00:00
abstract：:Growing evidence indicates that RNA G-quadruplexes have important roles in various processes such as transcription, translation, regulation of telomere length, and formation of telomeric heterochromatin. Investigation of RNA G-quadruplex structures associated with biological events is therefore essential to understand...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2017.156
更新日期：2018-04-01 00:00:00
