Towards a full quantitative description of yeast metabolism a systematic approach for estimating the kinetic parameters of isoenzymes under in vivo like conditions.

abstract：:We have described a method using polyvalent peptide display on filamentous phage that can be used to identify ligands that bind to G protein beta gamma subunits. Also described is how to construct phage that have known beta gamma binding sequences fused to the coat protein to allow a competition analysis to be perform...

journal_title：Methods in enzymology

authors： Smrcka AV,Scott JK

更新日期：2002-01-01 00:00:00

abstract：:G-protein-coupled receptors (GPCRs) form a superfamily of membrane proteins that play a crucial role in mediating physiological processes as well as pathogenesis of many critical diseases. They are one of the most successful drug targets, accounting for more than 30% of prescription drugs on the market today. Three-di...

journal_title：Methods in enzymology

authors： Vaidehi N,Pease JE,Horuk R

更新日期：2009-01-01 00:00:00

abstract：:ADP-ribosylation is a protein post-translational modification that is critically involved in a wide array of biological processes connected to cell stress responses. Enzymes known as poly-ADP-ribose polymerases (PARPs) catalyze the addition of the ADP-ribose units to amino acids with various side chain chemistries. In...

journal_title：Methods in enzymology

authors： Li P,Zhen Y,Yu Y

更新日期：2019-01-01 00:00:00

abstract：:5-FOA is an extremely useful reagent for the selection of Ura- cells amid a population of Ura+ cells. The selection is effective in transformation and recombination studies where loss of URA3+ is desired. A new plasmid shuffling procedure based on the 5-FOAR selection permits the recovery of conditional lethal mutatio...

journal_title：Methods in enzymology

authors： Boeke JD,Trueheart J,Natsoulis G,Fink GR

更新日期：1987-01-01 00:00:00

abstract：:Dicer is a multidomain ribonuclease that processes double-stranded RNAs (dsRNAs) to 21-nt small interfering RNAs (siRNAs) during RNA interference and excises microRNAs (miRNAs) from precursor hairpins. PAZ and PIWI domain (PPD) proteins, also involved in RNAi and miRNA function, are the best-characterized proteins kno...

journal_title：Methods in enzymology

authors： Kolb FA,Zhang H,Jaronczyk K,Tahbaz N,Hobman TC,Filipowicz W

更新日期：2005-01-01 00:00:00

abstract：:Prior to the development of 3D reconstruction, images were interpreted in terms of models made from simple units like ping-pong balls. Generally, people eye-balled the images and with other knowledge about its structure, such as the number of subunits, proposed models to account for the images. How was one to know if ...

journal_title：Methods in enzymology

authors： Derosier D

更新日期：2010-01-01 00:00:00

abstract：:RNA and DNA carry out diverse functions in biology including catalysis, splicing, gene regulation, and storage of genetic information. Interest has grown in understanding how nucleic acids perform such sophisticated functions given their limited molecular repertoire. RNA can fold into diverse shapes that often perturb...

journal_title：Methods in enzymology

authors： Thaplyal P,Bevilacqua PC

更新日期：2014-01-01 00:00:00

abstract：:Myoglobin has recently emerged as a versatile metalloprotein scaffold for the design of efficient and selective biocatalysts for abiological carbene transfer reactions, including asymmetric cyclopropanation reactions. Over the past few years, our group has explored several strategies to modulate the carbene transfer r...

journal_title：Methods in enzymology

authors： Carminati DM,Moore EJ,Fasan R

更新日期：2020-01-01 00:00:00

abstract：:Protein kinase C (PKC), a family of serine/threonine kinases, plays an important role in apoptosis. Several members of the PKC family act as substrates for caspases. In addition, PKCs can also regulate caspase activation and cell death by apoptosis. The cleavage of PKCs separates the regulatory domain from the catalyt...

journal_title：Methods in enzymology

authors： Basu A,Persaud SD,Sivaprasad U

更新日期：2008-01-01 00:00:00

abstract：:A genetic interaction occurs when the combination of two mutations leads to an unexpected phenotype. Screens for synthetic genetic interactions have been used extensively to identify genes whose products are functionally related. In particular, synthetic lethal genetic interactions often identify genes that buffer one...

journal_title：Methods in enzymology

authors： Baryshnikova A,Costanzo M,Dixon S,Vizeacoumar FJ,Myers CL,Andrews B,Boone C

更新日期：2010-01-01 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：:Carbohydrate microarrays are becoming a standard tool for glycobiologists to screen large numbers of sugars and elucidate the role of carbohydrates in biological systems. This article describes detailed methods to prepare and use microarrays containing synthetic oligosaccharides as well as a summary of the biological ...

journal_title：Methods in enzymology

authors： de Paz JL,Horlacher T,Seeberger PH

更新日期：2006-01-01 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：:Nanotechnology offers a new platform for therapeutic delivery of antiretrovirals to the central nervous system (CNS) where human immunodeficiency virus (HIV-1) is sequestered in patients with HIV-1-associated neurological disorders (HAND). HAND is a spectrum of neurocognitive disorders that continue to persist in HIV-...

journal_title：Methods in enzymology

authors： Mahajan SD,Law WC,Aalinkeel R,Reynolds J,Nair BB,Yong KT,Roy I,Prasad PN,Schwartz SA

更新日期：2012-01-01 00:00:00

abstract：:kappaB-Ras1 and kappaB-Ras2 are two small proteins that display similarity at the amino acid level to Ras-like small GTPases. Although little is known about the function of the kappaB-Ras proteins, they have been shown to interfere with activation of transcription factor NF-kappaB. They accomplish this by binding to I...

journal_title：Methods in enzymology

authors： Huxford T,Ghosh G

更新日期：2006-01-01 00:00:00

abstract：:ADP-ribosylation factor domain protein 1 (ARD1) is a multifunctional protein that belongs to the family of 20-kDa ARF proteins. The ARD1 gene encodes a 64-kDa protein with a structure comprising an 18-kDa ADP-ribosylation factor (ARF) domain at the C-terminus (amino acids 403-574), and a 46-kDa N-terminal domain (amin...

journal_title：Methods in enzymology

authors： Vichi A,Moss J,Vaughan M

更新日期：2005-01-01 00:00:00

abstract：:Polyacrylamide gel electrophoresis under native conditions (native PAGE) is a well-established and versatile method for probing nucleic acid conformation and nucleic acid-protein interactions. Native PAGE has been used to measure RNA folding equilibria and kinetics under a wide variety of conditions. Advantages of thi...

journal_title：Methods in enzymology

authors： Woodson SA,Koculi E

更新日期：2009-01-01 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：:Desmoplakin (DP) is an obligate component of desmosomes, where it links the desmosomal cadherin/plakoglobin/plakophilin assembly to intermediate filaments. DP contains a large amino-terminal domain (DPNT) that binds to the cadherin/plakoglobin/plakophilin complex, a central coiled-coil domain that dimerizes the molecu...

journal_title：Methods in enzymology

authors： Choi HJ,Weis WI

更新日期：2016-01-01 00:00:00

abstract：:Adenoviral vectors expressing vascular permeability factor/vascular endothelial growth factor (VPF/VEGF, VEGF-A(164)) offer a powerful method for elucidating the mechanisms of pathological angiogenesis and lymphangiogenesis and for evaluating the effectiveness of pro- and anti-angiogenesis therapies. When injected int...

journal_title：Methods in enzymology

authors： Nagy JA,Shih SC,Wong WH,Dvorak AM,Dvorak HF

更新日期：2008-01-01 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：:Autophagy is an extremely dynamic process that mediates the rapid degradation of intracellular components in response to different stress conditions. The autophagic response is executed by specific protein complexes, whose function is regulated by posttranslational modifications and interactions with positive and nega...

journal_title：Methods in enzymology

authors： Antonioli M,Ciccosanti F,Dengjel J,Fimia GM

更新日期：2017-01-01 00:00:00

abstract：:This chapter outlined the methods used to study intra- and extracellular modifications of apolipoproteins. These and other related studies have shown that several of the apolipoproteins undergo a series of intra- and extracellular modifications as follows: All apolipoproteins studied contain an 18-26 long signal pepti...

journal_title：Methods in enzymology

authors： Zannis VI,Karathanasis SK,Forbes GM,Breslow JL

更新日期：1986-01-01 00:00:00

abstract：:Intramembrane-cleaving proteases (I-CLiPs) are a group of membrane-associated proteases with a unique feature: they are believed to cleave their substrate within the hydrophobic lipid bilayer, even though peptide bond hydrolysis requires a water molecule. Escherichia coli RseP, which belongs to the S2P zinc metallopro...

journal_title：Methods in enzymology

authors： Hizukuri Y,Akiyama K,Akiyama Y

更新日期：2017-01-01 00:00:00

abstract：:Since the onset of the genomic era more than 1000 bacterial genomes have been sequenced and several fold more are expected to be completed in the near future. These genome sequences supply a wealth of information that can be exploited by statistical methods to gain significant insights into cellular processes. In Volu...

journal_title：Methods in enzymology

authors： Lunt B,Szurmant H,Procaccini A,Hoch JA,Hwa T,Weigt M

更新日期：2010-01-01 00:00:00

abstract：:The human cannabinoid receptor, CB1, has been difficult to purify in a functional form, hampering structural and biophysical studies. Here, we present our approaches for obtaining pure, detergent solubilized, functional CB1. We also discuss our site-directed fluorescence labeling (SDFL) methods for identifying differe...

journal_title：Methods in enzymology

authors： Fay JF,Farrens DL

更新日期：2017-01-01 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：: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：: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