Synthesis of bile acid coenzyme a thioesters in the amino acid conjugation of bile acids.

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：:The RNA-guided, sequence-specific endonuclease Cas9 has been widely adopted as genome engineering tool due to its efficiency and ease of use. Derived from the microbial CRISPR (clustered regularly interspaced short palindromic repeat) type II adaptive immune system, Cas9 has now been successfully engineered for genome...

journal_title：Methods in enzymology

authors： Trevino AE,Zhang F

更新日期：2014-01-01 00:00:00

abstract：:Reviewing the circumstances that have led to the first monoclonal antibody against the disease-associated form of PrP, we consider the availability of PrP knockout mice and recombinant PrP, as well as a reliable conformational screening protocol as being important prerequisites for a successful immunization approach. ...

journal_title：Methods in enzymology

authors： Korth C,Streit P,Oesch B

更新日期：1999-01-01 00:00:00

abstract：:The analysis of a reaction pathway by stopped-flow and chemical quench-flow methods provides the only means of directly measuring the rates of reaction at the active site of an enzyme. The interpretation of data from each technique is similar and involves solution of the differential equations describing the approach ...

journal_title：Methods in enzymology

authors： Johnson KA

更新日期：1986-01-01 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：:Vitamin K-dependent carboxylation, an essential posttranslational modification catalyzed by gamma-glutamyl carboxylase, is required for the biological functions of proteins that control blood coagulation, vascular calcification, bone metabolism, and other important physiological processes. Concomitant with carboxylati...

journal_title：Methods in enzymology

authors： Tie JK,Stafford DW

更新日期：2017-01-01 00:00:00

abstract：:The Ras-regulatory network is a loosely defined composition of numerous Ras family members and effector pathways that couple to critical cell-regulatory processes. Investigators are increasingly turning to RNAi-mediated inhibition of gene expression as an effective tool to help generate authentic portraits of Ras prot...

journal_title：Methods in enzymology

authors： Whitehurst AW,White MA

更新日期：2006-01-01 00:00:00

abstract：:The Duffy blood group antigen is a serpentine protein with seven transmembrane domains that is not coupled to G-proteins or other known intracellular effectors. In addition to erythrocytes, it is also expressed in endothelial cells and neurons. In recent years the Duffy antigen has received much attention because of i...

journal_title：Methods in enzymology

authors： Rot A,Horuk R

更新日期：2009-01-01 00:00:00

abstract：:Live cell microscopy using fluorescent proteins and small fluorescent probes is a well-established and essential tool for cell biology; however, there is a considerable need for noninvasive techniques able to study tissue and cell dynamics without the need to introduce chemical or genetically encoded probes. Coherent ...

journal_title：Methods in enzymology

authors： Pope I,Langbein W,Borri P,Watson P

更新日期：2012-01-01 00:00:00

abstract：:When yeast are faced with irreparable DNA damage, they will first arrest in G2/M, via the DNA damage checkpoint pathway, but will subsequently adapt to that arrest and resume division. Here, we summarize assays that we have used to examine checkpoint adaptation. Specifically, we discuss the merits of inducing DNA dama...

journal_title：Methods in enzymology

authors： Toczyski DP

更新日期：2006-01-01 00:00:00

abstract：:It has become increasingly clear that large RNA molecules, especially long noncoding RNAs, function in almost all gene regulatory processes (Cech & Steitz, 2014). Many large RNAs appear to be structural scaffolds for assembly of important RNA/protein complexes. However, the structures of most large cellular RNA molecu...

journal_title：Methods in enzymology

authors： Lin Y,May GE,Joel McManus C

更新日期：2015-01-01 00:00:00

abstract：:Assessment of protein nitration is commonly used as a footprint for the formation of reactive nitrogen species in vivo. However, one of the major disadvantages of measuring nitrotyrosine in proteins is that nitrated proteins are broken down at variable rates, and the resulting free nitrotyrosine is taken up by cells, ...

journal_title：Methods in enzymology

authors： Mani AR,Moore KP

更新日期：2005-01-01 00:00:00

abstract：:The target-of-rapamycin (TOR) pathway is a major regulator of cellular growth, division, and metabolism by coupling growth and nutrient cues to fundamental processes such as translation and transcription. In vertebrate models, disruptions affecting components of the TOR pathway result in cilia size abnormalities and c...

journal_title：Methods in enzymology

authors： Yuan S,Zhao L,Sun Z

更新日期：2013-01-01 00:00:00

abstract：:The Arc (anoxic redox control) two-component system (TCS) is a complex signal transduction system that plays an important role in regulating energy metabolism at the level of transcription in bacteria. This system comprises the ArcB protein, a hybrid membrane-associated sensor kinase, and the ArcA protein, a typical r...

journal_title：Methods in enzymology

authors： Alvarez AF,Georgellis D

更新日期：2010-01-01 00:00:00

abstract：:Apical caspases 8, 9, and 10 are only active as dimers. These dimers are unstable, and to characterize their activity they need to be maintained in vitro in a dimeric state. We provide updated methods for those looking to characterize various aspects of caspase function. We describe full methods for those looking to a...

journal_title：Methods in enzymology

authors： Mace PD,Riedl SJ,Salvesen GS

更新日期：2014-01-01 00:00:00

abstract：:The existence of an accurate force field (FF) model that reproduces the free-energy landscape is a key prerequisite for the simulation of biomineralization. Here, the stages in the development of such a model are discussed including the quality of the water model, the thermodynamics of polymorphism, and the free energ...

journal_title：Methods in enzymology

authors： Raiteri P,Demichelis R,Gale JD

更新日期：2013-01-01 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：:Riboswitches present a ubiquitous genetic regulatory mechanism for prokaryotes and have been found in HIV1, fungi, plants, and even H. sapiens. We present an overview of approaches to predict riboswitch aptamers and, more generally, RNA conformational switches. ...

journal_title：Methods in enzymology

authors： Clote P

更新日期：2015-01-01 00:00:00

abstract：:Accurate diagnosis of attentional disorders such as attention-deficit hyperactivity disorder (ADHD) is imperative because there are multiple negative psychosocial sequelae related to undiagnosed and untreated ADHD. Early and accurate detection can lead to effective intervention and prevention of negative sequelae. Unf...

journal_title：Methods in enzymology

authors： Robeva R,Penberthy JK

更新日期：2009-01-01 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：: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：:Classifications and characterizations of specific proteins, such as enzymes, not only allow us to understand biosynthetic and metabolic pathways but they also help to drive our understanding of protein structure and function. How those characterizations are evaluated, however, may change our interpretations and lead u...

journal_title：Methods in enzymology

authors： Bradshaw HB,Leishman E

更新日期：2017-01-01 00:00:00

abstract：:Heterodimeric complexes between individual members of the R7 subfamily of regulators of G-protein signaling proteins and the Gbeta5 isoform of the heterotrimeric G-protein beta subunit family are strongly expressed in the cell nucleus in neurons and brain, as well as in the cytoplasm and plasma membrane. Native and re...

journal_title：Methods in enzymology

authors： Simonds WF,Woodard GE,Zhang JH

更新日期：2004-01-01 00:00:00

abstract：:Enzymatic processes have great potential for applications in industrial biocatalysis, biosensing, and biomedical engineering. However, poor stability and difficulty in the reuse of enzymes still create challenges for their use in catalysis and other applications. In recent years, enzyme-incorporated metal-organic fram...

journal_title：Methods in enzymology

authors： Hou M,Ge J

更新日期：2017-01-01 00:00:00

abstract：:Recent advances have demonstrated the feasibility and robustness of chemical synthesis for the production of homogeneously glycosylated protein forms (glycoforms). By taking advantage of the unmatchable flexibility and precision provided by chemical synthesis, the quantitative effects of glycosylation were obtained us...

journal_title：Methods in enzymology

authors： Li Y,Tran AH,Danishefsky SJ,Tan Z

更新日期：2019-01-01 00:00:00

abstract：:Regulator of G-protein signaling (RGS) domains bind directly to GTP-bound Galpha subunits and accelerate their intrinsic GTPase activity by up to several thousandfold. The selectivity of RGS proteins for individual Galpha subunits has been illustrated. Thus, the expression of RGS proteins can be used to inhibit signal...

journal_title：Methods in enzymology

authors： Hains MD,Siderovski DP,Harden TK

更新日期：2004-01-01 00:00:00

abstract：:Gene therapy vectors based on murine retroviruses have now been in clinical trials for over 20 years. During that time, a variety of novel vector pseudotypes were developed in an effort to improve gene transfer. Lentiviral vectors are now in clinical trials and a similar evolution of vector technology is anticipated. ...

journal_title：Methods in enzymology

authors： Leath A,Cornetta K

更新日期：2012-01-01 00:00:00

abstract：:Death receptors are a subgroup of the tumor necrosis factor receptor superfamily (TNFRSF) and mediate activation of what is widely known as the "extrinsic" apoptosis pathway. TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) is one of the most recent death receptor ligands identified. The TRAIL receptor ...

journal_title：Methods in enzymology

authors： Harper N,MacFarlane M

更新日期：2008-01-01 00:00:00

abstract：:Cell migration plays a key role both in physiological conditions, such as tissue repair or embryonic development, and in pathological processes, including tumor metastasis. Understanding the mechanisms that allow cancer cells to invade tissues during metastasis requires studying their ability to migrate. While spectac...

journal_title：Methods in enzymology

authors： Vinot S,Anguille C,de Toledo M,Gadea G,Roux P

更新日期：2008-01-01 00:00:00

abstract：:Glycosaminoglycans (GAGs) are long unbranched polysaccharides, most of which are linked to a core protein to form proteoglycans. Depending on the nature of their backbone, one can discern galactosaminoglycans (chondroitin sulfate [CS] and dermatan sulfate [DS]) and glucosaminoglycans (heparan sulfate [HS], heparin, hy...

journal_title：Methods in enzymology

authors： Smits NC,Lensen JF,Wijnhoven TJ,Ten Dam GB,Jenniskens GJ,van Kuppevelt TH

更新日期：2006-01-01 00:00:00