How to operate a cryo-electron microscope.

abstract：:Rab7, a member of the Rab family of small G proteins, has been shown to regulate late endocytic traffic and lysosome biogenesis, but the exact roles and the mode of actions of Rab7 are still undetermined. Accumulating evidence suggests that each Rab protein has multiple target proteins and works together with them to ...

journal_title：Methods in enzymology

authors： Mizuno K,Sakane A,Sasaki T

更新日期：2005-01-01 00:00:00

abstract：:Formation of 5-methyluridine (ribothymidine) at position 54 of the T-psi loop of tRNA is catalyzed by site-specific tRNA methyltransferases (tRNA[uracil-54,C5]-MTases). In eukaryotes and many bacteria, the methyl donor for this reaction is generally S-adenosyl-L-methionine (S-AdoMet). However, in other bacteria, like ...

journal_title：Methods in enzymology

authors： Urbonavicius J,Brochier-Armanet C,Skouloubris S,Myllykallio H,Grosjean H

更新日期：2007-01-01 00:00:00

abstract：:Staphylococcus aureus ribonuclease III (Sa-RNase III) belongs to the enzyme family known to process double-stranded RNAs consisting of two turns of the RNA helix. Although the enzyme is thought to play a role in ribosomal RNA processing and gene regulation, the deletion of the rnc gene in S. aureus does not affect cel...

journal_title：Methods in enzymology

authors： Chevalier C,Huntzinger E,Fechter P,Boisset S,Vandenesch F,Romby P,Geissmann T

更新日期：2008-01-01 00:00:00

abstract：:Neurons are probably the most highly polarized cell type and typically develop a single axon and several dendrites. The establishment of a polarized morphology and the functional specialization of axonal and dendritic compartments are essential steps in the differentiation of neurons. Primary cultures of dissociated h...

journal_title：Methods in enzymology

authors： Schwamborn JC,Li Y,Püschel AW

更新日期：2006-01-01 00:00:00

abstract：:The method presented here is intended as a compromise between finding a good overall alignment and the time taken to do so. Many multiple alignment algorithms spend an excessively large amount of effort trying to find the best global alignment. This time is often ill spent because the results of the standard dynamic p...

journal_title：Methods in enzymology

authors： Taylor WR

更新日期：1990-01-01 00:00:00

abstract：:We have developed an array-based whole-genome genotyping (WGG) assay (Infinium) using our BeadChip platform that effectively enables unlimited multiplexing and unconstrained single nucleotide polymorphism (SNP) selection. A single tube whole-genome amplification reaction is used to amplify the genome, and loci of inte...

journal_title：Methods in enzymology

authors： Gunderson KL,Steemers FJ,Ren H,Ng P,Zhou L,Tsan C,Chang W,Bullis D,Musmacker J,King C,Lebruska LL,Barker D,Oliphant A,Kuhn KM,Shen R

更新日期：2006-01-01 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

abstract：:The coat protein complex II (COPII) coat is responsible for direct capture of membrane cargo proteins and for the physical deformation of the endoplasmic reticulum (ER) membrane that drives the transport vesicle formation. The use of an in vitro reconstitution system comprising purified components is desirable for stu...

journal_title：Methods in enzymology

authors： Sato K,Nakano A

更新日期：2005-01-01 00:00:00

abstract：:A central goal of biology is to understand how transcription factors target and regulate specific genes and networks to control cell fate and function. An equally important goal of synthetic biology, chemical biology, and personalized medicine is to devise molecules that can regulate genes and networks in a programmab...

journal_title：Methods in enzymology

authors： Tietjen JR,Donato LJ,Bhimisaria D,Ansari AZ

更新日期：2011-01-01 00:00:00

abstract：:Membrane-bound pyrophosphatases couple the hydrolysis of inorganic pyrophosphate to the pumping of ions (sodium or protons) across a membrane in order to generate an electrochemical gradient. This class of membrane protein is widely conserved across plants, fungi, archaea, and bacteria, but absent in multicellular ani...

journal_title：Methods in enzymology

authors： Harborne SPD,Strauss J,Turku A,Watson MA,Tuma R,Harris SA,Goldman A

更新日期：2018-01-01 00:00:00

abstract：:Cancer has become the leading cause of death in the developed world and has remained one of the most difficult diseases to treat. One of the difficulties in treating cancer is that conventional chemotherapies often have unacceptable toxicities toward normal cells at the doses required to kill tumor cells. Thus, the de...

journal_title：Methods in enzymology

authors： Cantor JR,Panayiotou V,Agnello G,Georgiou G,Stone EM

更新日期：2012-01-01 00:00:00

abstract：:Inducible expression systems represent the founding technology for the emergence of synthetic biology in mammalian cells. The core molecules in these systems are bacterial regulator proteins that bind to or dissociate from a cognate DNA operator sequence in response to an exogenous stimulus like a small-molecule induc...

journal_title：Methods in enzymology

authors： Karlsson M,Weber W,Fussenegger M

更新日期：2011-01-01 00:00:00

abstract：:Ion channels open and close in response to diverse stimuli, and the molecular events underlying these processes are extensively modulated by ligands of both endogenous and exogenous origin. In the past decade, high-resolution structures of several channel types have been solved, providing unprecedented details of the ...

journal_title：Methods in enzymology

authors： Chakrapani S

更新日期：2015-01-01 00:00:00

abstract：:Enzymology is approaching an era where many problems can benefit from computational studies. While ample challenges remain in quantitatively predicting behavior for many enzyme systems, the insights that often come from computations are an important asset for the enzymology community. Here we provide a primer for enzy...

journal_title：Methods in enzymology

authors： Roston D,Cui Q

更新日期：2016-01-01 00:00:00

abstract：:Nonsense-mediated mRNA decay (NMD) is activated by exon-junction complexes (EJCs) that are located downstream of the termination codon of the substrate mRNAs. This situation can be imitated by tethering components of the EJC to the 3' untranslated region (3' UTR) of a reporter mRNA. Here we describe the detailed use o...

journal_title：Methods in enzymology

authors： Gehring NH,Hentze MW,Kulozik AE

更新日期：2008-01-01 00:00:00

abstract：:Actinomycete bacteria of the genus Streptomyces are major producers of bioactive compounds for the biotechnology industry. They are the source of most clinically used antibiotics, as well as of several widely used drugs against common diseases, including cancer . Genome sequencing has revealed that the potential of St...

journal_title：Methods in enzymology

authors： Medema MH,Breitling R,Takano E

更新日期：2011-01-01 00:00:00

abstract：:By far the highest resolution of all separation techniques for intact proteins in a single analytical run continues to be by the combination of isoelectric focusing (IEF) and SDS-PAGE, originally introduced by O'Farrell [(1975). J. Biol. Chem.250, 4007-4021]. This analytical platform has seen a number of significant a...

journal_title：Methods in enzymology

authors： Friedman DB,Hoving S,Westermeier R

更新日期：2009-01-01 00:00:00

abstract：:The fact that chemical exchange processes occur at rates that cover a broad range and produce readily detectable effects on the spectrum is one of the attractive features of high-resolution NMR. The description of these line shapes in the presence of spin-spin coupling requires the density matrix theory which is rathe...

journal_title：Methods in enzymology

authors： Rao BD

更新日期：1989-01-01 00:00:00

abstract：:Throughout the history of single-particle electron microscopy (EM), automated technologies have seen varying degrees of emphasis and development, usually depending upon the contemporary demands of the field. We are currently faced with increasingly sophisticated devices for specimen preparation, vast increases in the ...

journal_title：Methods in enzymology

authors： Lyumkis D,Moeller A,Cheng A,Herold A,Hou E,Irving C,Jacovetty EL,Lau PW,Mulder AM,Pulokas J,Quispe JD,Voss NR,Potter CS,Carragher B

更新日期：2010-01-01 00:00:00

abstract：:Inositol phosphates (InsPs) are an important group of eukaryotic messengers and mediate a wide range of processes. To elucidate the biological functions of these molecules, robust techniques to characterize inositol phosphate metabolism at the cellular level are highly sought after. This chapter provides a detailed pr...

journal_title：Methods in enzymology

authors： Puschmann R,Harmel RK,Fiedler D

更新日期：2020-01-01 00:00:00

abstract：:The field of therapeutic application of the A₃ adenosine receptor (A₃AR) antagonists represents a rapidly growing and intense area of research in the adenosine field. Even if there are currently no A₃AR antagonists in clinical phases, in light of the plethora of biological effects attributed to A₃ARs, substantial effo...

journal_title：Methods in enzymology

authors： Paoletta S,Federico S,Spalluto G,Moro S

更新日期：2010-01-01 00:00:00

abstract：:Methane production and consumption in anaerobic marine sediments is catalyzed by a series of reversible tetrahydromethanopterin (H(4)MPT)-linked C1 transfer reactions. Although many of these reactions are conserved between one-carbon compound utilizing microorganisms, two remain diagnostic for archaeal methane metabol...

journal_title：Methods in enzymology

authors： Hallam SJ,Pagé AP,Constan L,Song YC,Norbeck AD,Brewer H,Pasa-Tolic L

更新日期：2011-01-01 00:00:00

abstract：:The above methodology has been used to prepare a variety of PE-immunotoxins. These reagents are potent cytotoxic agents for cells in culture that bind the appropriate antibody and nontoxic when cells do not bind the antibody. PE-immunotoxins can be used to select for mutant cultured cells which lack the target of the ...

journal_title：Methods in enzymology

authors： FitzGerald DJ

更新日期：1987-01-01 00:00:00

abstract：:After cell lysis, the most often used second step in a protein purification procedure is some sort of a rapid, bulk precipitation step. This is commonly accomplished by altering the solvent conditions and taking advantage of the changes in solubility of your protein of interest relative to those of many of the other p...

journal_title：Methods in enzymology

authors： Burgess RR

更新日期：2009-01-01 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：:Receptor ensemble docking (RED) is an effective strategy to account for receptor flexibility in the course of a docking-based virtual screening campaign. Such an approach can be applied when multiple crystal structures of a receptor have been solved, but it can also be applied when only a single crystal structure is a...

journal_title：Methods in enzymology

authors： Vilar S,Costanzi S

更新日期：2013-01-01 00:00:00

abstract：:Gene regulatory grids (GRGs) encompass the space of all the possible transcription factor (TF)-target gene interactions that regulate gene expression, with gene regulatory networks (GRNs) representing a temporal and spatial manifestation of a portion of the GRG, essential for the specification of gene expression. Thus...

journal_title：Methods in enzymology

authors： Yang F,Ouma WZ,Li W,Doseff AI,Grotewold E

更新日期：2016-01-01 00:00:00

abstract：:This study designed, synthesized, and characterized a number of new heterobifunctional photocleavable cross-linking reagents that may be used to photomodulate the activity of proteins or to prepare caged fluorescent dyes. Biomolecules or fluorophores caged via a thiol group with the BNBASE reagent can be covalently li...

journal_title：Methods in enzymology

authors： Marriott G,Ottl J

更新日期：1998-01-01 00:00:00

abstract：:The study of hormone secretion by anterior pituitary cells is complicated by the presence of multiple cell types. For unambiguous interpretation of data it is necessary to identify the cells from which measurements are made. We have described a reliable experimental approach involving the identification of cultured ce...

journal_title：Methods in enzymology

authors： Croxton TL,Armstrong WM,Ben-Jonathan N

更新日期：1989-01-01 00:00:00

abstract：:In recent years, sphingolipid metabolites ceramide, sphingosine, and sphingosine-1-phosphate have emerged as important second messengers in addition to their role as precursors of biomembrane components. The investigation of these sphingolipid metabolites requires the development of new, more sensitive methods for ass...

journal_title：Methods in enzymology

authors： Spassieva S,Bielawski J,Anelli V,Obeid LM

更新日期：2007-01-01 00:00:00