Intra- and extracellular modifications of apolipoproteins.

abstract：:Genetically encodable RNA devices that directly detect small molecules in the cellular environment are of increasing interest for a variety of applications including live cell imaging and synthetic biology. Riboswitches are naturally occurring sensors of intracellular metabolites, primarily found in the bacterial mRNA...

journal_title：Methods in enzymology

authors： Trausch JJ,Batey RT

更新日期：2015-01-01 00:00:00

abstract：:Pathogen-host interactions are mediated in part by secreted microbial proteins capable of exploiting host cells for their survival. Several of these manipulations involve, but are not limited to, suppression of defense responses, alterations in host vesicular trafficking, and manipulation of gene expression. The deliv...

journal_title：Methods in enzymology

authors： Clark HR,Hayes TA,Kale SD

更新日期：2014-01-01 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：:Genetic modifier screens offer a powerful, indeed a uniquely powerful tool for the analysis and identification of elements capable of modulating specific cellular functions in development. Here, we describe the methodology that allowed us to explore the genetic circuitry that affects a Notch mutant phenotype caused by...

journal_title：Methods in enzymology

authors： Hori K,Sen A,Artavanis-Tsakonas S

更新日期：2014-01-01 00:00:00

abstract：:IF-liposomes are formed by a unique process that involves fusing small liposomes into interdigitated lipid sheets, using either ethanol or hydrostatic pressure. The interdigitation-fusion method requires liposome formulations with lipids that form the L beta I phase. Preparing ethanol-induced IF-liposomes is simple an...

journal_title：Methods in enzymology

authors： Ahl PL,Perkins WR

更新日期：2003-01-01 00:00:00

abstract：:Organ-specific delivery of biofunctional agents is thought to enhance their activity and to reduce their side effects. Liposomes have been used as drug carriers in cancer chemotherapy, since they accumulate passively in tumor tissues due to an enhanced permeability and retention (EPR) effect. In addition, modification...

journal_title：Methods in enzymology

authors： Asai T,Oku N

更新日期：2005-01-01 00:00:00

abstract：:Site-directed mutagenesis is a PCR-based method to mutate specified nucleotides of a sequence within a plasmid vector. This technique allows one to study the relative importance of a particular amino acid for protein structure and function. Typical mutations are designed to disrupt or map protein-protein interactions,...

journal_title：Methods in enzymology

authors： Bachman J

更新日期：2013-01-01 00:00:00

abstract：:Endocytosis and autophagy are key vesicular pathways involved in degradation and recycling of cellular material. Both degradative pathways finally fuse with lysosome but are indeed interconnected at several levels. In particular, the fusion between endosomes and autophagosomes can generate intermediate vesicles named ...

journal_title：Methods in enzymology

authors： Manil-Ségalen M,Culetto E,Legouis R,Lefebvre C

更新日期：2014-01-01 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：:T lymphocytes express on their surface a heterodimeric αβ receptor, called the T cell receptor (TCR), which recognizes foreign antigens. Unlike antibodies, the recognition requires both an antigenic peptide epitope and a protein encoded by the major histocompatibility complex (MHC). In contrast to conventional antibod...

journal_title：Methods in enzymology

authors： Stone JD,Chervin AS,Aggen DH,Kranz DM

更新日期：2012-01-01 00:00:00

abstract：:Superoxide dismutases (SODs; EC 1.15.1.1) are a group of metalloenzymes which are essential to protect cells under aerobic conditions. In biological systems, it has been reported that SODs and other proteins are susceptible to be attacked by peroxynitrite (ONOO(-)) which can be originated from the reaction of nitric o...

journal_title：Methods in enzymology

authors： Larrainzar E,Urarte E,Auzmendi I,Ariz I,Arrese-Igor C,González EM,Moran JF

更新日期：2008-01-01 00:00:00

abstract：:The Microsealed Drug Delivery (MDD) system was developed to achieve several objectives: to maximize the bioavailability of a therapeutic agent in a target tissue; to minimize the adverse side effects of a therapeutic agent or its metabolites; to optimize the onset, rate, and duration of drug delivery; to maintain the...

journal_title：Methods in enzymology

authors： Chien YW

更新日期：1985-01-01 00:00:00

abstract：:Since the early days of cell volume regulation research, the role of actin cytoskeleton organization and rearrangement has attracted specific interest. Rapid modifications in actin dynamics and architecture have been described. They were shown to regulate cell volume changes, as well as regulatory volume decrease in a...

journal_title：Methods in enzymology

authors： Papakonstanti EA,Stournaras C

更新日期：2007-01-01 00:00:00

abstract：:Nedd8 is a ubiquitin-like molecule that is highly conserved in eukaryotes. Similar to ubiquitin, Nedd8 attaches to target proteins through an enzymatic cascade composed of Nedd8-specific E1 (activating)- and E2 (conjugating)-enzymes. The E1 for Nedd8 is a heterodimer of APP-BP1 and Uba3, while the E2 is Ubc12. The mos...

journal_title：Methods in enzymology

authors： Chiba T

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：:The Ras gene family has been implicated in the development of many human epithelial cancers. Mutations in K-ras or its downstream mediator BRAF have been detected in about two thirds of low-grade serous carcinomas and borderline serous tumors; mutations in K-ras are also often present in benign and invasive mucinous o...

journal_title：Methods in enzymology

authors： Rosen DG,Yang G,Bast RC Jr,Liu J

更新日期：2006-01-01 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：:Significant progress has been made in discovering and cloning a host of proteins, including a range of glycoproteins. The availability of their predicted amino acid sequences provides useful information, including potential N-linked glycosylation sites. However, only a limited number of protein structures have been so...

journal_title：Methods in enzymology

authors： Yen TY,Macher BA

更新日期：2006-01-01 00:00:00

abstract：:The methodologies described in this chapter allow the reproducible preparation of native high-molecular-weight synthetase complexes of leuRL. These complexes have the ability to preferentially utilize extracellular leucine immediately upon transport and are likely the forms of the enzyme most important in the utilizat...

journal_title：Methods in enzymology

authors： Hampel A,Tritz R

更新日期：1988-01-01 00:00:00

abstract：:As for other GPCRs, the oligomerization of glycoprotein hormone receptors (GPHRs) appears as critical event for receptor function. By means of modern techniques based on the BRET or FRET principle, GPHR oligomerization has been reported to explain several physiological and pathological conditions. In particular, the p...

journal_title：Methods in enzymology

authors： Bonomi M,Persani L

更新日期：2013-01-01 00:00:00

abstract：:Cells belonging to the monocyte/macrophage lineage are in general highly resistant to peroxynitrite. Resistance is not dependent on the scavenging of peroxynitrite itself, or of other secondary reactive species, but is rather associated with the prompt activation of a survival signaling leading to the prevention of to...

journal_title：Methods in enzymology

authors： Cantoni O,Tommasini I,Cerioni L

更新日期：2008-01-01 00:00:00

abstract：:Biominerals have complex and heterogeneous architectures, hence diffraction experiments with spatial resolutions between 500 nm and 10 μm are extremely useful to characterize them. X-ray beams in this size range are now routinely produced at many synchrotrons. This chapter provides a review of the different hard X-ray...

journal_title：Methods in enzymology

authors： Tamura N,Gilbert PU

更新日期：2013-01-01 00:00:00

abstract：:The RNA degradosome is a bacterial protein machine devoted to RNA degradation and processing. In Escherichia coli, it is typically composed of the endoribonuclease RNase E, which also serves as a scaffold for the other components: the exoribonuclease PNPase, the RNA helicase RhlB, and enolase. The variable presence of...

journal_title：Methods in enzymology

authors： Mauri P,Dehò G

更新日期：2008-01-01 00:00:00

abstract：:Nonnative protein aggregation has been classically treated as an amorphous process occurring by colloidal coagulation kinetics and proceeding to an essentially irreversible endpoint often ascribed to a chaotic tangle of unfolded chains. However, some nonnative aggregates, particularly amyloid fibrils, exhibit ordered ...

journal_title：Methods in enzymology

authors： O'Nuallain B,Thakur AK,Williams AD,Bhattacharyya AM,Chen S,Thiagarajan G,Wetzel R

更新日期：2006-01-01 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：:In vivo genome manipulation through site-directed mutagenesis and chromosome rearrangements has been hindered by the difficulty in achieving high frequencies of targeting and the intensive labor required to create altered genomes that do not contain any heterologous sequence. Here we describe our approach, referred to...

journal_title：Methods in enzymology

authors： Storici F,Resnick MA

更新日期：2006-01-01 00:00:00

abstract：:Dynamin I is a large GTPase enzyme required in membrane constriction and fission during multiple forms of endocytosis. The first method described here is for the rapid purification of native dynamin from peripheral membrane extracts of sheep brain using ammonium sulfate precipitation and affinity purification on recom...

journal_title：Methods in enzymology

authors： Quan A,Robinson PJ

更新日期：2005-01-01 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：:Nonribosomal peptides are natural products typically of bacterial and fungal origin. These highly complex molecules display a broad spectrum of biological activities, and have been exploited for the development of immunosuppressant, antibiotic, anticancer, and other therapeutic agents. The nonribosomal peptides are as...

journal_title：Methods in enzymology

authors： Wilkinson B,Micklefield J

更新日期：2009-01-01 00:00:00

abstract：:A simple and convenient method is described to determine primary deuterium kinetic isotope effects (1°DKIEs) on reactions where the hydron incorporated into the reaction product is derived from solvent water. The 1°DKIE may be obtained by 1H NMR analyses as the ratio of the yields of H- and D-labeled products from a r...

journal_title：Methods in enzymology

authors： Amyes TL,Richard JP

更新日期：2017-01-01 00:00:00