Use of laser microdissection in complex tissue.

abstract：:Protein-protein interactions (PPIs) are universal to life and their study and understanding is critical to drug discovery and bioengineering efforts. Historically, X-ray crystallography, isothermal titration calorimetry and other biophysical methods have been used to study PPIs, but can be costly and are low throughpu...

journal_title：Methods in enzymology

authors： Charov K,Burkart MD

更新日期：2020-01-01 00:00:00

abstract：:Adiponectin is a circulating bioactive hormone secreted by adipocytes as oligomers ranging in size from 90 kDa trimers and 180 kDa hexamers to larger high molecular weight oligomers that may reach 18- or 36-mers in size. While total circulating adiponectin levels correlate well with metabolic health, it is the relativ...

journal_title：Methods in enzymology

authors： Rutkowski JM,Scherer PE

更新日期：2014-01-01 00:00:00

abstract：:The gasdermin (GSDM) family consists of gasdermin A (GSDMA), B (GSDMB), C (GSDMC), D (GSDMD), E or DNFA5 (GSDME), and DFNB59 in human. Expressed in the skin, gastrointestinal tract, and various immune cells, GSDMs mediate homeostasis and inflammation upon activation by caspases and unknown proteases. In particular, GS...

journal_title：Methods in enzymology

authors： Xia S,Ruan J,Wu H

更新日期：2019-01-01 00:00:00

abstract：:Osmoregulation is the active control of the cellular water balance and encompasses homeostatic mechanisms crucial for life. The osmoregulatory system in the yeast Saccharomyces cerevisiae is particularly well understood. Key to yeast osmoregulation is the production and accumulation of the compatible solute glycerol, ...

journal_title：Methods in enzymology

authors： Hohmann S,Krantz M,Nordlander B

更新日期：2007-01-01 00:00:00

abstract：:Signal transduction by vascular endothelial growth factors (VEGFs) through their cognate VEGF receptor tyrosine kinases follows the consensus scheme for receptor tyrosine kinases. Thus, binding of ligand induces receptor dimerization and activation of the tyrosine kinase through transphosphorylation between receptor m...

journal_title：Methods in enzymology

authors： Li X,Claesson-Welsh L,Shibuya M

更新日期：2008-01-01 00:00:00

abstract：:In eukaryotes, determination of the subcellular location of a novel protein encoded in genomic or transcriptomic data provides useful clues as to its possible function. However, experimental localization studies are expensive and time-consuming. As a result, accurate in silico prediction of subcellular localization fr...

journal_title：Methods in enzymology

authors： Gaston D,Tsaousis AD,Roger AJ

更新日期：2009-01-01 00:00:00

abstract：:Model building studies may be used to supplement structurally low resolution experimental data with detailed three-dimensional hypothetical atomic models. Because of the strong relation between structure and function in biological molecules such models may give a consistent, integral view of a wealth of experimental d...

journal_title：Methods in enzymology

authors： von Kitzing E

更新日期：1992-01-01 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：:A fundamental concept of phosphorylation-mediated signaling is the precise switching between discrete functional conformations. According to the traditional view, phosphorylation induces a new, active conformation. In this chapter, a series of NMR experiments performed on a response regulator are described that challe...

journal_title：Methods in enzymology

authors： Gardino AK,Kern D

更新日期：2007-01-01 00:00:00

abstract：:Reference standard materials (RSMs) exist for a variety of biologics including vaccines but are not readily available for gene therapy vectors. To date, a recombinant adeno-associated virus serotype 2 RSM (rAAV2 RSM) has been produced and characterized and was made available to the scientific community in 2010. In add...

journal_title：Methods in enzymology

authors： Moullier P,Snyder RO

更新日期：2012-01-01 00:00:00

abstract：:Enveloped virus particles select their lipid-protein components and egress by budding from the host cell membranes. The matrix protein of many enveloped viruses has been proposed as a crucial element for viral budding; however, molecular mechanisms behind membrane remodeling by the matrix protein are yet to be unravel...

journal_title：Methods in enzymology

authors： Shnyrova AV,Zimmerberg J

更新日期：2009-01-01 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：: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：: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：:Virtually all eukaryotic cell functions and signaling pathways are regulated by protein phosphorylation. The Rad53 kinase plays crucial roles in the DNA damage response in Saccharomyces cerevisiae and is widely used as a surrogate marker for DNA damage checkpoint activation by diverse genotoxic agents. Most currently ...

journal_title：Methods in enzymology

authors： Hoch NC,Chen ES,Tsai MD,Heierhorst J

更新日期：2017-01-01 00:00:00

abstract：:Once the appropriate site has been selected for the attachment of GFP to the sarcomeric protein, it is quite remarkable that the large size of the GFP molecule does not appear to interfere with the localization of the fluorescent sarcomeric proteins into the sarcomeric regions of the myofibrils. A similar approach usi...

journal_title：Methods in enzymology

authors： Dabiri GA,Ayoob JC,Turnacioglu KK,Sanger JM,Sanger JW

更新日期：1999-01-01 00:00:00

abstract：:PRA2 was found to interact with the ER-localized protein VAMP-associated protein of 33 kDa or VAP-33 by a yeast two-hybrid screen. We describe here the purification of PRA2 and VAP-33 as well as an in vitro pull-down procedure to verify the interaction. PRA2 was found to form a large sodium dodecyl sulfate (SDS)-insol...

journal_title：Methods in enzymology

authors： Gougeon PY,Ngsee JK

更新日期：2005-01-01 00:00:00

abstract：:Nonribosomal peptide biosynthesis is a complex enzymatic assembly responsible for producing a great diversity of bioactive peptide natural products. Due to the recurring arrangement of catalytic domains within these machineries, great interest has been shown in reengineering these pathways to produce novel, designer p...

journal_title：Methods in enzymology

authors： Greule A,Charkoudian LK,Cryle MJ

更新日期：2019-01-01 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：:Mathematical computer-assisted simulations of episodic hormone, substrate, or metabolite release can be accomplished using explicit algebra and high-speed microprocessors. Such simulations are useful to describe the behavior of single or multiple pulse generators, as well as the expected output of these systems. Simul...

journal_title：Methods in enzymology

authors： Veldhuis JD,Johnson ML

更新日期：1994-01-01 00:00:00

abstract：:The diverse structures of polyketide natural products are reflected by the equally diverse polyketide biosynthetic enzymes, namely polyketide synthases (PKSs). Three major classes of PKSs are known-noniterative type I PKSs, iterative type II PKSs and acyl carrier protein-independent type III PKSs, each of which consis...

journal_title：Methods in enzymology

authors： Cheng YQ,Coughlin JM,Lim SK,Shen B

更新日期：2009-01-01 00:00:00

abstract：:We describe the methodologies for the design of artificial enzymes with genetically encoded unnatural amino acids. Genetically encoded unnatural amino acids offer great promise for constructing artificial enzymes with novel activities. In our studies, the designs of artificial enzyme were divided into two steps. First...

journal_title：Methods in enzymology

authors： Hu C,Wang J

更新日期：2016-01-01 00:00:00

abstract：:The protein kinase C (PKC) family of isoenzymes may be a crucial player in transducing H2O2-induced signaling in a wide variety of physiological and pathophysiological processes. PKCs contain unique structural features that make them highly susceptible to oxidative modification. Depending on the site of oxidation and ...

journal_title：Methods in enzymology

authors： Gopalakrishna R,McNeill TH,Elhiani AA,Gundimeda U

更新日期：2013-01-01 00:00:00

abstract：:Histone chaperones-key actors in the dynamic organization of chromatin-interact with the various histone variants to ensure their transfer in and out of chromatin. In vitro chromatin assembly assays and isolation of protein complexes using tagged histone variants provided first clues concerning their binding specifici...

journal_title：Methods in enzymology

authors： Clément C,Vassias I,Ray-Gallet D,Almouzni G

更新日期：2016-01-01 00:00:00

abstract：:Precise investigation and manipulation of gene function often require modulation in a controlled and dynamic manner. In this chapter, we describe the methods to apply HIT systems for drug inducible transcriptional activation or simultaneous activation and genome editing in human cells. Together with those for editing,...

journal_title：Methods in enzymology

authors： Zhao C,Wei S,Wang Y

更新日期：2019-01-01 00:00:00

abstract：:A significant proportion of RNA decay in eukaryotes takes place in the nucleus. In addition to targeting nongenic RNAs such as products of processing and spurious transcription events, nuclear RNA decay also removes aberrant transcripts from regular genes. Moreover, a small list of genes utilizes nuclear RNA turnover ...

journal_title：Methods in enzymology

authors： Schmid M,Rougemaille M,Libri D,Jensen TH

更新日期：2008-01-01 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：:Over the past 19 years, we have developed a novel myxoma virus-derived anti-inflammatory serine protease inhibitor, termed a serpin, as a new class of immunomodulatory therapeutic. This review will describe the initial identification of viral serpins with anti-inflammatory potential, beginning with preclinical analysi...

journal_title：Methods in enzymology

authors： Chen H,Zheng D,Davids J,Bartee MY,Dai E,Liu L,Petrov L,Macaulay C,Thoburn R,Sobel E,Moyer R,McFadden G,Lucas A

更新日期：2011-01-01 00:00:00

abstract：:Mass spectrometry (MS) has gradually replaced classical methods as a major tool in protein sequencing and characterization. However, the sample preparation repertoire has not changed very much; it has just been adjusted to the needs of the new analytical method. In this chapter frequently used in-solution enzymatic di...

journal_title：Methods in enzymology

authors： Medzihradszky KF

更新日期：2005-01-01 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