Medicinal chemistry inspired fragment-based drug discovery.

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：:A number of different analytical techniques are now available for the isolation of apoA-I, apoA-II, and apoA-IV. The choice of a particular technique is dependent on the instrumentation available, and the quantity of isolated apolipoprotein required. The isolation and characterization of the separate isoforms and the ...

journal_title：Methods in enzymology

authors： Brewer HB Jr,Ronan R,Meng M,Bishop C

更新日期：1986-01-01 00:00:00

abstract：:Acidic organelles and vesicles, such as endosomes, lysosomes, autophagosomes, trans-Golgi network, and synaptic vesicles, are known to play important roles in a broad range of cellular events. To facilitate studying these multifunctional systems, we describe here an acid-brightening fluorescent protein (abFP), which f...

journal_title：Methods in enzymology

authors： Wang N,Wang L

更新日期：2020-01-01 00:00:00

abstract：:Although the function of histone deacetylases (HDACs) have primarily been associated with influencing transcription through chromatin remodeling, the capacity of these enzymes to interface with a diverse array of biologic processes by modulating a growing list of nonhistone substrates has gained recent attention. Rece...

journal_title：Methods in enzymology

authors： Kahali S,Sarcar B,Chinnaiyan P

更新日期：2011-01-01 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 combined use of rotating-frame relaxation methods, temperature-dependent measurements of line shapes and magnetization transfer experiments allows in favorable cases the examination in some detail of exchange processes that occur on the millisecond time scale. It is possible to determine not only the rate constant...

journal_title：Methods in enzymology

authors： Lane AN,Lefèvre JF

更新日期：1994-01-01 00:00:00

abstract：:Neurofilament biology is important to understanding structural properties of axons, such as establishment of axonal diameter by radial growth. In order to study the function of neurofilaments, a series of genetically modified mice have been generated. Here, we describe a brief history of genetic modifications used to ...

journal_title：Methods in enzymology

authors： Jones MR,Villalón E,Garcia ML

更新日期：2016-01-01 00:00:00

abstract：:Ubiquitin (Ub)-mediated protein degradation is a key cellular defense mechanism that detects and eliminates defective proteins. A major intracellular site of protein quality control degradation is the endoplasmic reticulum (ER), hence the term ER-associated degradation, or endoplasmic reticulum-associated degradation ...

journal_title：Methods in enzymology

authors： Mashahreh B,Reiss Y,Wiener R,Ravid T

更新日期：2019-01-01 00:00:00

abstract：:Immunogenic cell death (ICD), induced by certain anticancer chemotherapeutics, leads to the emission of danger associated molecular patterns (DAMP) by cancer cells, which facilitates the attraction, activation and maturation of dendritic cells (DC) as well as the subsequent priming of effector T cells. In this context...

journal_title：Methods in enzymology

authors： Cerrato G,Liu P,Martins I,Kepp O,Kroemer G

更新日期：2020-01-01 00:00:00

abstract：:Tumor cells acquire distinct genetic characteristics as a means to survive and proliferate indefinitely. Changes in the genetic code can also translate in changes at the protein level, therefore creating a distinguishable signature unique for tumor cells, and absent in normal tissues. The presence of discernable moiet...

journal_title：Methods in enzymology

authors： Battaglia S

更新日期：2020-01-01 00:00:00

abstract：:This chapter discusses experimental methods and protocols for the purification and preliminary characterization of DNA polymerases that are specialized for the replicative bypass (translesion DNA synthesis) of base or other types of DNA damage that typically arrest high-fidelity DNA synthesis, with particular emphasis...

journal_title：Methods in enzymology

authors： Fischhaber PL,McDaniel LD,Friedberg EC

更新日期：2006-01-01 00:00:00

abstract：:We have used a shortened construct form of the CYP26A1 gene promoter, in a promoter-less vector with either luciferase (known as E4) or a red fluorescent protein, RFP (known as E4.2) as the reporter gene and examined their responses to retinoids in transfected HepG2 and HEK293T cells. The promoter responded linearly t...

journal_title：Methods in enzymology

authors： Zolfaghari R,Mattie FJ,Wei CH,Chisholm DR,Whiting A,Ross AC

更新日期：2020-01-01 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：:Nucleic acid aptamers find widespread use as targeting and sensing agents in nature and biotechnology. Their ability to bind an extensive range of molecular targets, including small molecules, proteins, and ions, with high affinity and specificity enables their use in diverse diagnostic, therapeutic, imaging, and gene...

journal_title：Methods in enzymology

authors： Chang AL,McKeague M,Smolke CD

更新日期：2014-01-01 00:00:00

abstract：:Genetically encoded fluorescent sensors are essential tools in modern biological research, and recent advances in fluorescent proteins (FPs) have expanded the scope of sensor design and implementation. In this review we compare different sensor platforms, including Förster resonance energy transfer (FRET) sensors, flu...

journal_title：Methods in enzymology

authors： Sanford L,Palmer A

更新日期：2017-01-01 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：:Autophagy is a catabolic process that is crucial for cellular homeostasis and adaptive response to changing environments. Importantly, autophagy has been shown to be induced in many longevity-associated scenarios and to be required to maintain lifespan extension. Notably, autophagy is a highly conserved cellular proce...

journal_title：Methods in enzymology

authors： Kainz K,Tadic J,Zimmermann A,Pendl T,Carmona-Gutierrez D,Ruckenstuhl C,Eisenberg T,Madeo F

更新日期：2017-01-01 00:00:00

abstract：:Many methods have been developed for the cloning of PCR products. These methods include blunt-end cloning, TA cloning, and using restriction sites incorporated into the PCR primers. The restrictionless cloning technique allows efficient directional cloning of PCR products into any cloning site within a vector regardle...

journal_title：Methods in enzymology

authors： Algire MA

更新日期：2013-01-01 00:00:00

abstract：:The combination of PCR amplification and chemiluminescent detection of PCR products provides a highly sensitive system for the quantitation of DNA and RNA. The broad dynamic range of the chemiluminescent detection assay simplifies the selection of cycling and concentration parameters critical to harnessing the quantit...

journal_title：Methods in enzymology

authors： Martin CS,Voyta JC,Bronstein I

更新日期：2000-01-01 00:00:00

abstract：:Lysosomes are specialized organelles for protein recycling and as such are involved in the terminal steps of autophagy. However, it has become evident that lysosomes also play an important role in the progression of apoptosis. This latter function seems to be dependent on lysosomal proteases, which need to be released...

journal_title：Methods in enzymology

authors： Ivanova S,Repnik U,Bojic L,Petelin A,Turk V,Turk B

更新日期：2008-01-01 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：:Chloroplast genomes of land plants and algae contain generally between 100 and 150 genes. These genes are involved in plastid gene expression and photosynthesis and in various other tasks. The function of some chloroplast genes is still unknown and some of them appear to be essential for growth and survival. Repressib...

journal_title：Methods in enzymology

authors： Ramundo S,Rochaix JD

更新日期：2015-01-01 00:00:00

abstract：:A synthetic ligand regulable system for gene transfer and expression has been developed in our laboratory based on mechanistic studies of steriod hormone receptor and transcriptional regulation. This gene switch system possesses most of the important features that are required for application of the system in biologic...

journal_title：Methods in enzymology

authors： Ye X,Schillinger K,Burcin MM,Tsai SY,O'Malley BW

更新日期：2002-01-01 00:00:00

abstract：:Cryptococcus neoformans is a yeastlike fungus that causes a lethal meningoencephalitis in a broad spectrum of immunocompromised patients and has become the most common cause of meningitis due to AIDS-related infections in Africa. Key to the development of new agents to control and prevent this infection is the identif...

journal_title：Methods in enzymology

authors： Hu G,Gibbons J,Williamson PR

更新日期：2008-01-01 00:00:00

abstract：:Intracellular calcium (Ca2+) is involved in the regulation of a variety of biological functions in cancer cells, including growth inhibition, tumor invasiveness, and drug resistance. To gain insight into the possible role played by Ca2+ in the development of drug resistance in breast cancer, we performed a comparative...

journal_title：Methods in enzymology

authors： DeBernardi MA,Brooker G

更新日期：2006-01-01 00:00:00

abstract：:The lung is composed of two major anatomically distinct regions-the conducting airways and gas-exchanging airspaces. From a cell biology standpoint, the conducting airways can be further divided into two major compartments, the tracheobronchial and bronchiolar airways, while the alveolar regions of the lung make up th...

journal_title：Methods in enzymology

authors： Liu X,Driskell RR,Engelhardt JF

更新日期：2006-01-01 00:00:00

abstract：:In actively growing cells, the rate of translation initiation is relatively rapid. As a result, multiple ribosomes simultaneously engaged in translation become spaced along single mRNA molecules. These structures, termed polysomes, can be separated from ribosomal subunits and single ribosomes because they migrate fast...

journal_title：Methods in enzymology

authors： Qin D,Fredrick K

更新日期：2013-01-01 00:00:00

abstract：:Human embryonic stem cells hold great promise in furthering our treatment of disease and increasing our understanding of early development. This chapter describes protocols for the derivation and maintenance of human embryonic stem cells. In addition, it summarizes briefly several alternative methods for the culture o...

journal_title：Methods in enzymology

authors： Akutsu H,Cowan CA,Melton D

更新日期：2006-01-01 00:00:00

abstract：:Quantitative real-time polymerase chain reaction (qRT-PCR) is a flexible and scalable method for analyzing transcript abundance that can be used at a single gene or high-throughput (>100 genes) level. Information obtained from this technique can be used as an indicator of potential regulation of glycosylation at the t...

journal_title：Methods in enzymology

authors： Nairn AV,dela Rosa M,Moremen KW

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