Bacterial expression and purification of vertebrate odorant-binding proteins.

abstract：:Unfolded protein response (UPR) is a major reaction to intrinsic stress of eukaryotic organisms and is also related to environmental stress reactions. Among yeasts, stress regulation has mainly been investigated in Saccharomyces cerevisiae, while other species with biotechnological or medical interest are less well un...

journal_title：Methods in enzymology

authors： Dragosits M,Mattanovich D,Gasser B

更新日期：2011-01-01 00:00:00

abstract：:This chapter presents the application of capillary electrophoresis coupled to electrospray mass spectrometry (CE-ES-MS) for the analysis of complex bacterial lipopolysaccharides (LPS) from pathogenic strains of Haemophilus influenzae and Neisseria meningitidis. A discussion is included of the development of electropho...

journal_title：Methods in enzymology

authors： Li J,Martin A,Cox AD,Moxon ER,Richards JC,Thibault P

更新日期：2005-01-01 00:00:00

abstract：:Aliphatic carbon-halogen (C-X) bonds are prevalent in modern pharmaceuticals and bioactive natural products. Three distinct chemical strategies are known in Nature to generate these structural motifs. The first is via the nucleophilic substitution at a prefunctionalized electrophilic carbon center with a halide anion ...

journal_title：Methods in enzymology

authors： Liu X

更新日期：2018-01-01 00:00:00

abstract：:Membrane fission, which divides membrane surfaces into separate compartments, is essential to diverse cellular processes including membrane trafficking and cell division. Quantitative assays are needed to elucidate the physical mechanisms by which proteins drive membrane fission. Toward this goal, several experimental...

journal_title：Methods in enzymology

authors： Snead WT,Stachowiak JC

更新日期：2018-01-01 00:00:00

abstract：:There is a great need for practicing scientists to volunteer their time and expertise in the K-12th grade science classroom. We have found that bioluminescence is a fun and exciting way to teach basic science concepts and is an excellent tool for the volunteering scientist. We have had very positive reactions from bot...

journal_title：Methods in enzymology

authors： Hammer M,Andrade JD

更新日期：2000-01-01 00:00:00

abstract：:Intrinsically disordered proteins (IDPs) sample structurally diverse ensembles. Characterizing the underlying distributions of conformations is a key step toward understanding the structural and functional properties of IDPs. One increasingly popular method for obtaining quantitative information on intramolecular dist...

journal_title：Methods in enzymology

authors： Holmstrom ED,Holla A,Zheng W,Nettels D,Best RB,Schuler B

更新日期：2018-01-01 00:00:00

abstract：:Enzymes in the α-d-phosphohexomutase (PHM) superfamily catalyze a multistep reaction, entailing two successive phosphoryl transfers. Key to this reaction is a conserved phosphoserine in the active site, which serves alternately as a phosphoryl donor and acceptor during the catalytic cycle. In addition to its role in t...

journal_title：Methods in enzymology

authors： Stiers KM,Beamer LJ

更新日期：2018-01-01 00:00:00

abstract：:Recent advances in functional genomics have enabled genome-wide genetic studies in mammalian cells. These include the establishment of high-throughput transfection and viral propagation methodologies, the production of large-scale cDNA and siRNA libraries, and the development of sensitive assay detection processes and...

journal_title：Methods in enzymology

authors： Rines DR,Tu B,Miraglia L,Welch GL,Zhang J,Hull MV,Orth AP,Chanda SK

更新日期：2006-01-01 00:00:00

abstract：:We describe here the components and uses of two image intensifier systems. The SIT camera system is convenient, relatively inexpensive, readily adaptable to most microscopes, and reliable. It suffers from lack of resolution at high gain levels and the inability to extend its sensitivity by accumulating an image over t...

journal_title：Methods in enzymology

authors： Willingham MC,Pastan IH

更新日期：1983-01-01 00:00:00

abstract：:Oligomerization is a frequently encountered physical characteristic of biological molecules that occurs for a wide number of transcription factors, ion channels, oxygen-carrying macromolecules such as hemocyanin and enzymes. On the other hand, unwanted protein oligomerization can lead to the formation of pathogenic st...

journal_title：Methods in enzymology

authors： Barranco-Medina S,Dietz KJ

更新日期：2009-01-01 00:00:00

abstract：:This protocol describes Coomassie brilliant blue staining, one of the most common methods of detecting proteins in polyacrylamide gels (PAGE). ...

journal_title：Methods in enzymology

authors： Brunelle JL,Green R

更新日期：2014-01-01 00:00:00

abstract：:Delivery of drugs and molecules to eukaryote animal and plant cells can be enhanced when mediated by mesoporous silica nanoparticle (MSN) materials. This chapter describes, in detail, the synthesis of several different MSN materials used for the transport of various hydrophilic and hydrophobic drug molecules, neurotra...

journal_title：Methods in enzymology

authors： Fang IJ,Trewyn BG

更新日期：2012-01-01 00:00:00

abstract：:Linkers play essential roles in the engineering of fusion proteins, and have been extensively demonstrated to affect protein properties such as expression level, solubility, and biological functions. For linker design and optimization, one of the key factors is the flexibility or rigidity of linkers, which describes t...

journal_title：Methods in enzymology

authors： Huang Z,Zhang C,Xing XH

更新日期：2021-01-01 00:00:00

abstract：:The breaking of RNA strands by 2'-O-transphosphorylation is a ubiquitous reaction in biology, and enzymes that catalyze this reaction play key roles in RNA metabolism. The mechanisms of 2'-O-transphosphorylation in solution are relatively well studied, but complex and can involve different transition states depending ...

journal_title：Methods in enzymology

authors： Harris ME,York DM,Piccirilli JA,Anderson VE

更新日期：2017-01-01 00:00:00

abstract：:Helical symmetry is commonly used for building macromolecular assemblies. Helical symmetry is naturally present in viruses and cytoskeletal filaments and also occurs during crystallization of isolated proteins, such as Ca-ATPase and the nicotinic acetyl choline receptor. Structure determination of helical assemblies b...

journal_title：Methods in enzymology

authors： Diaz R,Rice WJ,Stokes DL

更新日期：2010-01-01 00:00:00

abstract：:There is currently no known genetic disease linked to prolactin (PRL) or its receptor (PRLR) in humans. Recently, we identified three missense variants of the PRLR in patients presenting with breast tumors. Two of them (named PRLR(I146L) and PRLR(I76V)) had been reported earlier, but failed to draw much attention beca...

journal_title：Methods in enzymology

authors： Goffin V,Bogorad RL,Touraine P

更新日期：2010-01-01 00:00:00

abstract：:Given the increasing evidence of oxidative stress in AD brain and studies from different perspectives that appear to show a converging, central role for A beta in the pathogenesis and etiology of AD, insight into A beta-associated free radical oxidative stress will likely lead to a greater understanding of AD and, pot...

journal_title：Methods in enzymology

authors： Butterfield DA,Yatin SM,Varadarajan S,Koppal T

更新日期：1999-01-01 00:00:00

abstract：:In mice with lupus nephritis qualitative changes in anti-DNA antibodies occur, such as IgG switch and increased cationic charge, to render these antibodies pathogenic. Pathogenic anti-DNA idiotypes can be encoded by genes of a normal mouse strain such as SWR, where they remain dormant. When the normal mice are crossed...

journal_title：Methods in enzymology

authors： Datta SK

更新日期：1988-01-01 00:00:00

abstract：:Patient-derived xenografts are a useful tool in cancer immunology, as they allow researchers to study human cancers in vivo when starting with a relatively small amount of human tumor tissue. These models make it possible to study tumor cell-intrinsic changes that occur in response to external stimuli including cytoki...

journal_title：Methods in enzymology

authors： Cardenas JJ,Robles-Oteiza C,Politi K

更新日期：2020-01-01 00:00:00

abstract：:Rab38 is a low-molecular-weight G-protein highly expressed in melanocytes of the skin and alveolar type II cells in the lung. A point mutation in the postulated GTP/GDP-interacting domain of Rab38 has been identified as the genetic lesion responsible for oculocutaneous albinism (OCA) in chocolate (cht) mice. Another p...

journal_title：Methods in enzymology

authors： Osanai K,Voelker DR

更新日期：2008-01-01 00:00:00

abstract：:RNA folds during transcription in the cell. Compared to most in vitro studies where the focus is generally on Mg(2+)-initiated refolding of fully synthesized transcripts, cotranscriptional RNA folding studies better replicate how RNA folds in a cellular environment. Unique aspects of cotranscriptional folding include ...

journal_title：Methods in enzymology

authors： Wong TN,Pan T

更新日期：2009-01-01 00:00:00

abstract：:High content, phenotypic screens offer a powerful approach to systems biology at the cellular level. The approach employs cells carrying fluorescently labeled molecules or organelles in 384- or 1536-well microplates, and an automated confocal screening microscope for capturing images from each well. Although some spec...

journal_title：Methods in enzymology

authors： Kepiro M,Varkuti BH,Davis RL

更新日期：2018-01-01 00:00:00

abstract：:A neuron is able to seamlessly respond to a number of signals, in a timely and specific manner. This process, of integrating multiple inputs, relays on the orchestration of intracellular events by signaling networks. The inherent complexity of signaling networks has made computational modeling a useful approach to und...

journal_title：Methods in enzymology

authors： Wenderski WC,Neves SR

更新日期：2012-01-01 00:00:00

abstract：:Enveloped virus particles select their lipid-protein components and egress by budding from the host cell membranes. The matrix protein of many enveloped viruses has been proposed as a crucial element for viral budding; however, molecular mechanisms behind membrane remodeling by the matrix protein are yet to be unravel...

journal_title：Methods in enzymology

authors： Shnyrova AV,Zimmerberg J

更新日期：2009-01-01 00:00:00

abstract：:Peptides corresponding to the calmodulin-binding domain of MLCK and other target enzymes promise to be useful tools in the study of calmodulin action. Not only will they help to elucidate the molecular interactions of calmodulin with specific target enzymes, but they should also prove to be useful calmodulin antagonis...

journal_title：Methods in enzymology

authors： Blumenthal DK,Krebs EG

更新日期：1987-01-01 00:00:00

abstract：:Mass spectrometry (MS) has gradually replaced classical methods as a major tool in protein sequencing and characterization. However, the sample preparation repertoire has not changed very much; it has just been adjusted to the needs of the new analytical method. In this chapter frequently used in-solution enzymatic di...

journal_title：Methods in enzymology

authors： Medzihradszky KF

更新日期：2005-01-01 00:00:00

abstract：:Polarized epithelial cells are characterized by distinct plasma membrane domains and asymmetrical distribution of cell surface proteins and lipids. In vertebrates, tight junctions act as a fence between the apical and basolateral domains. Although many of the key components of the polarity machinery have been identifi...

journal_title：Methods in enzymology

authors： Chen X,Macara IG

更新日期：2006-01-01 00:00:00

abstract：:The functional properties of proteins are decided not only by their relatively rigid overall structures, but even more importantly, by their dynamic properties. In a protein, some regions of structure exhibit highly correlated or anti-correlated motions with others, some are highly dynamic but uncorrelated, while othe...

journal_title：Methods in enzymology

authors： Yu H,Dalby PA

更新日期：2020-01-01 00:00:00

abstract：:Epitope tagging has provided a useful experimental strategy with widespread applicability. The ample variety of epitope tag systems that have been put to use to date provide a collection of attributes relevant to virtually any experimental system. As a consequence, epitope tagging will continue to be a valuable tool f...

journal_title：Methods in enzymology

authors： Fritze CE,Anderson TR

更新日期：2000-01-01 00:00:00

abstract：:Acetyl CoA:arylamine N-acetyltransferase (NAT; E.C. 2.3.1.5) enzymes play a key role in the metabolic activation of aromatic amine and nitroaromatic mutagens to electrophilic reactive intermediates. We have developed a system in which the activation of mutagens by recombinant human NAT2, expressed in Escherichia coli,...

journal_title：Methods in enzymology

authors： Savulescu MR,Mushtaq A,Josephy PD

更新日期：2005-01-01 00:00:00