Solving serpin crystal structures.

Abstract:

:Essentially the same steps are required to solve the crystal structure of a serpin as for any other protein: produce and purify protein, grow crystals, collect diffraction data, find estimates of the phase angles, and then refine and validate the structure. For the phasing step, experimental phasing methods involving heavy atom soaks were required for the first few structures, but with the large number of serpin structures now available, molecular replacement has become the method of choice. Two things are special about serpins. First, because of the central role of conformational change in serpin mechanism, it is advisable to consider a variety of molecular replacement models in different conformations and then to allow for rigid-body motions in the initial refinement steps. Second, probably owing to the flexibility of serpins, the average serpin crystal is significantly less well ordered than the average crystal of another protein, which increases the difficulty of solving and refining their structures.

journal_name

Methods Enzymol

journal_title

Methods in enzymology

authors

Read RJ,Zhou A,Stein PE

doi

10.1016/B978-0-12-385950-1.00004-3

subject

Has Abstract

pub_date

2011-01-01 00:00:00

pages

49-61

eissn

0076-6879

issn

1557-7988

pii

B978-0-12-385950-1.00004-3

journal_volume

501

pub_type

杂志文章
  • Advances in phylogeny reconstruction from gene order and content data.

    abstract::Genomes can be viewed in terms of their gene content and the order in which the genes appear along each chromosome. Evolutionary events that affect the gene order or content are "rare genomic events" (rarer than events that affect the composition of the nucleotide sequences) and have been advocated by systematists for...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/S0076-6879(05)95035-0

    authors: Moret BM,Warnow T

    更新日期:2005-01-01 00:00:00

  • Studying RNA-RNA and RNA-protein interactions by isothermal titration calorimetry.

    abstract::Isothermal Titration Calorimetry (ITC) provides a sensitive and accurate means by which to study the thermodynamics of RNA folding, RNA binding to small molecules, and RNA-protein interactions. The advent of extremely sensitive instrumentation and the increasing availability of ITC in shared facilities have made it in...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/S0076-6879(09)68019-8

    authors: Feig AL

    更新日期:2009-01-01 00:00:00

  • Preparation of metagenomic libraries from naturally occurring marine viruses.

    abstract::Microbes are now well recognized as major drivers of the biogeochemical cycling that fuels the Earth, and their viruses (phages) are known to be abundant and important in microbial mortality, horizontal gene transfer, and modulating microbial metabolic output. Investigation of environmental phages has been frustrated ...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/B978-0-12-407863-5.00008-3

    authors: Solonenko SA,Sullivan MB

    更新日期:2013-01-01 00:00:00

  • Analysis of autophagy during infections of Cryptococcus neoformans.

    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

    pub_type: 杂志文章

    doi:10.1016/S0076-6879(08)03222-9

    authors: Hu G,Gibbons J,Williamson PR

    更新日期:2008-01-01 00:00:00

  • Studying antibody-antigen interactions with fluorescence fluctuation spectroscopy.

    abstract::Antibodies are excellent binding proteins that have found numerous applications in biological research, biotechnology, and medicine. Characterization of their ligand binding properties has long been, and continues to be, the focus of many researchers. Antibodies are also perfect test systems which can be used for the ...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/B978-0-12-405539-1.00005-1

    authors: Tetin SY,Ruan Q,Skinner JP

    更新日期:2013-01-01 00:00:00

  • Is this protein ubiquitinated?

    abstract::Covalent modification of proteins with ubiquitin plays an important role in a wide array of cellular processes (Hershko and Ciechanover, 1998; Pickart, 2004). For this reason an increasing number of investigators in diverse research fields are confronted with the question whether their favorite proteins are ubiquitina...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/S0076-6879(05)99016-2

    authors: Kaiser P,Tagwerker C

    更新日期:2005-01-01 00:00:00

  • Screening and characterizing human NAT2 variants.

    abstract::Acetyl CoA:arylamine N-acetyltransferase (NAT; E.C. 2.3.1.5) enzymes play a key role in the metabolic activation of aromatic amine and nitroaromatic mutagens to electrophilic reactive intermediates. We have developed a system in which the activation of mutagens by recombinant human NAT2, expressed in Escherichia coli,...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/S0076-6879(05)00011-X

    authors: Savulescu MR,Mushtaq A,Josephy PD

    更新日期:2005-01-01 00:00:00

  • Actin cytoskeleton architecture and signaling in osmosensing.

    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

    pub_type: 杂志文章

    doi:10.1016/S0076-6879(07)28012-7

    authors: Papakonstanti EA,Stournaras C

    更新日期:2007-01-01 00:00:00

  • Strategies in the Design and Use of Synthetic "Internal Glycan" Vaccines.

    abstract::The relative structural conservation of "internal glycans" in the cell walls of pathogens suggests that they might as target epitopes less prone to variation and hence with greater potential universality as vaccine targets. Examples of such glycans include the inner core sugars of lipopolysaccharides in Gram-negative ...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/bs.mie.2017.06.008

    authors: Lakshminarayanan A,Vijayakrishnan B,Bayley H,Davis BG

    更新日期:2017-01-01 00:00:00

  • Deuteration in Biological Neutron Reflectometry.

    abstract::Neutron reflectometry (NR) is uniquely positioned in structural biology, because of its ability to characterize biomolecular interfacial architectures like lipid membranes and membrane-associated proteins nondestructively and in their native environment. Mimicking biological processes, samples can be manipulated and t...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/bs.mie.2015.05.019

    authors: Heinrich F

    更新日期:2016-01-01 00:00:00

  • Harnessing RNAi for analyses of Ras signaling and transformation.

    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

    pub_type: 杂志文章

    doi:10.1016/S0076-6879(05)07022-9

    authors: Whitehurst AW,White MA

    更新日期:2006-01-01 00:00:00

  • Transport of a caspase inhibitor across the blood-brain barrier by chitosan nanoparticles.

    abstract::The current treatment of neurological and psychiatric diseases is far beyond being satisfactory. In addition to highly complex disease mechanisms, the blood-brain barrier (BBB) also remains as a challenge by limiting the delivery of the majority of currently available therapeutics to the central nervous system. Severa...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/B978-0-12-391860-4.00013-6

    authors: Yemişci M,Gürsoy-Özdemir Y,Caban S,Bodur E,Capan Y,Dalkara T

    更新日期:2012-01-01 00:00:00

  • Epitope tagging and protein surveillance.

    abstract::The epitope tagging approach offers advantages of economy, universality, and precision over the use of antibodies raised directly against a protein of interest. The latter strategy promises a potentially greater diversity of reagents and obviates the need to modify the protein, but it may not yield sufficiently high-a...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/0076-6879(91)94038-e

    authors: Kolodziej PA,Young RA

    更新日期:1991-01-01 00:00:00

  • Electrochemical biosensing with odorant binding proteins.

    abstract::The development of sensors that mimic the natural smell sensing mechanism and selectively recognizes the odorants remains highly challenging. Electrochemical based sensing approaches aiming at monitoring molecular recognition events between surface receptors and analytes in solution or in the gas phase, are one possib...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/bs.mie.2020.04.071

    authors: Szunerits S,Boukherroub R,Vasilescu A

    更新日期:2020-01-01 00:00:00

  • Membrane protein expression and analysis in yeast.

    abstract::This protocol describes the expression and analysis of membrane proteins produced in yeast, as illustrated with Yarrowia lipolytica and Pichia pastoris. Step by step, we explain how to generate a yeast strain expressing the membrane protein of interest, how to prepare a membrane protein sample from yeast, and how to a...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/bs.mie.2014.12.010

    authors: Claes K,Guerfal M,Callewaert N

    更新日期:2015-01-01 00:00:00

  • A Comparison of Two-Hybrid Approaches for Detecting Protein-Protein Interactions.

    abstract::Two-hybrid systems are one of the most popular, preferred, cost effective, and scalable in vivo genetic approaches for screening protein-protein interactions. A number of variants of yeast and bacterial two-hybrid systems exist, rendering them ideal for modern, flexible proteomics-driven studies. For mapping protein i...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/bs.mie.2016.10.020

    authors: Mehla J,Caufield JH,Sakhawalkar N,Uetz P

    更新日期:2017-01-01 00:00:00

  • Imaging of live malaria blood stage parasites.

    abstract::Life cell imaging is a tool for cell biology that has provided invaluable insights into many dynamic processes such as cell division, morphogenesis, or endo- and exocytosis. While observing cells by time-lapse imaging is a standard procedure in many systems, this technique was until recently not available for blood st...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/B978-0-12-391856-7.00029-9

    authors: Grüring C,Spielmann T

    更新日期:2012-01-01 00:00:00

  • Continuous cultivation and thermodynamic aspects of niche definition in the nitrogen cycle.

    abstract::The study of model organisms in pure culture has provided detailed information about the physiology and biochemistry of nitrification and related processes. Metagenomic sequencing of environmental samples is providing information to what extent this understanding also applies to natural microbial communities. Here, we...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/B978-0-12-381294-0.00002-X

    authors: Müller S,Strous M

    更新日期:2011-01-01 00:00:00

  • Transgenic models to understand hypoxia-inducible factor function.

    abstract::The hypoxia-inducible factor (HIF) is a heterodimeric basic helix-loop-helix (bHLH) transcription factor that controls the mammalian cellular transcriptional response to low oxygen tension by up-regulating genes including glycolytic enzymes and angiogenic factors, such as the vascular endothelial growth factor (VEGF)....

    journal_title:Methods in enzymology

    pub_type: 杂志文章,评审

    doi:10.1016/S0076-6879(07)35005-2

    authors: Doedens A,Johnson RS

    更新日期:2007-01-01 00:00:00

  • Peripheral clocks and the regulation of cardiovascular and metabolic function.

    abstract::Circadian rhythms generated by cell autonomous biological clocks allow for the appropriate temporal synchronization of physiology and behavior, optimizing the efficiency of biological systems. Circadian oscillators and functions have been uncovered in both central and peripheral tissues. This article describes methodo...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/S0076-6879(05)93027-9

    authors: Rudic RD,Curtis AM,Cheng Y,FitzGerald G

    更新日期:2005-01-01 00:00:00

  • Crystallization and X-ray structure determination of an RNA-dependent hexameric helicase.

    abstract::Hexameric helicases couple the energy of ATP hydrolysis to processive movement along nucleic acids and are critical components of cells and many viruses. Molecular motion derives from ATP hydrolysis at up to six distinct catalytic centers, which is coupled to the coordinated action of translocation loops in the center...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/B978-0-12-396546-2.00008-5

    authors: Thomsen ND,Berger JM

    更新日期:2012-01-01 00:00:00

  • Expression, purification, and assay of cytosolic (catalytic) domains of membrane-bound mammalian adenylyl cyclases.

    abstract::The identification and isolation of the soluble catalytic domains of adenylyl cyclase have provided investigators with useful reagents for the study of these enzymes. They have permitted detailed mechanistic investigation of the actions of forskolin, Gs alpha, and the inhibitory G protein, Gi alpha. They have served a...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/s0076-6879(02)45012-4

    authors: Hatley ME,Gilman AG,Sunahara RK

    更新日期:2002-01-01 00:00:00

  • Sedimentation Velocity Analysis of Large Oligomeric Chromatin Complexes Using Interference Detection.

    abstract::Sedimentation velocity experiments measure the transport of molecules in solution under centrifugal force. Here, we describe a method for monitoring the sedimentation of very large biological molecular assemblies using the interference optical systems of the analytical ultracentrifuge. The mass, partial-specific volum...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/bs.mie.2015.05.007

    authors: Rogge RA,Hansen JC

    更新日期:2015-01-01 00:00:00

  • Assessment of IFNγ responsiveness in patient-derived xenografts.

    abstract::Patient-derived xenografts are a useful tool in cancer immunology, as they allow researchers to study human cancers in vivo when starting with a relatively small amount of human tumor tissue. These models make it possible to study tumor cell-intrinsic changes that occur in response to external stimuli including cytoki...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/bs.mie.2019.10.027

    authors: Cardenas JJ,Robles-Oteiza C,Politi K

    更新日期:2020-01-01 00:00:00

  • Site-directed mutagenesis.

    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

    pub_type: 杂志文章

    doi:10.1016/B978-0-12-418687-3.00019-7

    authors: Bachman J

    更新日期:2013-01-01 00:00:00

  • Rapid kinetic analysis of mechanochemical adenosinetriphosphatases.

    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

    pub_type: 杂志文章

    doi:10.1016/0076-6879(86)34129-6

    authors: Johnson KA

    更新日期:1986-01-01 00:00:00

  • Applications of Dissolution-DNP for NMR Screening.

    abstract::Experimental screening for protein-ligand interactions is a central task in drug discovery. Nuclear magnetic resonance (NMR) spectroscopy enables the determination of binding affinities, as well as the measurement of structural and dynamic parameters governing the interaction. With traditional liquid-state NMR relying...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/bs.mie.2018.08.016

    authors: Kim Y,Hilty C

    更新日期:2019-01-01 00:00:00

  • Analysis of metabolically labeled inositol phosphate messengers by NMR.

    abstract::Inositol phosphates (InsPs) are an important group of eukaryotic messengers and mediate a wide range of processes. To elucidate the biological functions of these molecules, robust techniques to characterize inositol phosphate metabolism at the cellular level are highly sought after. This chapter provides a detailed pr...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/bs.mie.2020.04.035

    authors: Puschmann R,Harmel RK,Fiedler D

    更新日期:2020-01-01 00:00:00

  • Chapter 4 - Reconstitution of membrane budding with unilamellar vesicles.

    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

    pub_type: 杂志文章

    doi:10.1016/S0076-6879(09)64004-0

    authors: Shnyrova AV,Zimmerberg J

    更新日期:2009-01-01 00:00:00

  • Chapter 4. Analyzing the regulation of antibiotic production in streptomycetes.

    abstract::This chapter outlines the approaches and techniques that can be used to analyze the regulation of antibiotic production in streptomycetes. It describes how to isolate antibiotic nonproducing and overproducing mutants by UV, nitrosoguanidine (NTG), transposon, and insertion mutagenesis, and then how to use those mutant...

    journal_title:Methods in enzymology

    pub_type: 杂志文章

    doi:10.1016/S0076-6879(09)04804-6

    authors: Bibb M,Hesketh A

    更新日期:2009-01-01 00:00:00