rdf-syntax-grammar
mods:
  schema location: http://www.loc.gov/mods/v3 http://www.loc.gov/standards/mods/v3/mods-3-2.xsd
  title info:
    title: G Protein-Coupled Receptors
    part number: Part A
  name:
    name part: Shukla, Arun K.
    role:
      role term: editor.
  name:
    name part: ScienceDirect (Online service)
  type of resource: text
  genre: bibliography
  genre: Electronic books.
  origin info:
    place:
      place term: mau
    date issued: 2017
    copyright date: 2017
    edition: Second edition.
    issuance: monographic
  language:
    language term: eng
  physical description:
    form: remote
    extent: 1 online resource (xiii, 245 pages) : color illustrations.
  abstract: "The largest family of cell surface receptors in our body, referred to as G protein-coupled receptors, plays central roles in the onset and regulation of many different physiological processes. The cascade starts with binding of the "stimulus" (i.e., an activating ligand; agonist) to its cognate GPCR, followed by receptor activation, effector coupling and initiation of downstream signaling responses. Subsequently, the receptors are internalized as a mechanism of blunting the signaling response arising from the surface. Afterwards, the receptors are either recycled back to the plasma membrane to get ready for a new signaling cycle, or degraded via proteosomal and lysosomal mechanisms. In the latter case, newly synthesized receptors occupy the membrane, thus making the cell receptive again for the incoming stimulus. Development of precise and sensitive methods to measure the activation and signaling responses of GPCRs has been at the center stage of GPCR research. As a result, a number of assays have been designed, validated, and utilized to assess the ligand binding, effector coupling and receptor trafficking in cellular context. In addition, several approaches have also been developed and optimized to study ligand?receptor and receptor?effector interactions in vitro. Furthermore, direct computational approaches guided by the recent surge of structural data on GPCRs are also being developed rapidly to better understand the conformational dynamics and effector interactions of GPCRs. In this volume of Methods in Cell Biology entitled "Measuring GPCR Activation and Signaling", we have compiled a set of assays that are commonly used in many laboratories across the world to assess ligand-induced receptor activation, signaling, and regulation. We have covered cell based assays to record the interaction and activation of heterotrimeric G proteins with activated receptors, generation of second messengers (such as cAMP and Ca++), trafficking of activated receptors by confocal…
  table of contents: 3.3. Transfection of Cells With the G[gamma] Translocation Sensor Using the Cationic Polymer, PEI3.3.1. Preparation of PEI; 3.3.2. Transfection; 3.3.3. Other methods for transfection; 3.4. Live-Cell Time-Lapse Imaging; 3.4.1. Examination of optical activation of light-sensing GPCRs (Opsins); 3.5. Image Analysis; 3.5.1. Manual processing; 3.5.2. Automated/Thresholding processing; 3.6. Stable Cell Line Generation; References; Chapter 2: GloSensor assay for discovery of GPCR-selective ligands; 1. Introduction; 2. cAMP Measurement Assays; 2.1. Fluorescence-Based cAMP Measuring Assays
  table of contents: Front Cover; G Protein-Coupled Receptors Part A; Copyright; Contents; Contributors; Preface; Chapter 1: Measurement of GPCR-G protein activity in living cells; 1. Introduction; 2. Materials; 2.1. Generations of Sensors; 2.2. Cell Culture; 2.3. Transfection of Cells; 2.4. Stable Cell Line Generation; 2.5. Live-Cell Imaging; 2.6. Fluorescence Imaging Plate Reader Assay; 3. Methods; 3.1. Generations of Sensors Using Gibson Assembly Protocol; 3.2. Cell Culture; 3.2.1. HeLa Cell-Specific Growth Media Preparation; 3.2.2. Passaging cells
  table of contents: 2.1.1. ALPHAscreen (Amplified Luminescent Proximity Homogenous Assay)2.1.2. DELFIA (Dissociation-Enhanced Lanthanide Fluorescence Immunoassay); 2.1.3. HTRF (Homogenous Time Resolved Fluorescence); 2.1.4. Fluorescence polarization assay; 2.2. Luminescence-Based cAMP Measuring Assays; 2.2.1. CRE-luciferase reporter gene assay; 2.2.2. GloSensor assay; 3. Reagents and Consumables; 4. Methods; 4.1. Cell Culture Maintenance and Trypsinization; 4.2. Transient Transfection of HEK293T Cells; 4.3. Preparation of Transfected Cell Plate; 4.4. Test Compound Dilution for GloSensor Assay
  table of contents: 4.5. Test Compound Addition to the Cell Plate4.6. Compound Addition to Screen Ligands for GS-Coupled Receptors; 4.7. Compound Addition to Screen Ligands for Gi-Coupled Receptors; 5. Data Analysis; 5.1. Data Analysis for Agonist Activity; 5.2. Data Analysis for Antagonist Activity; 6. Results and Data Interpretation; 7. Final Comments; Acknowledgments; References; Chapter 3: cAMP assays in GPCR drug discovery; 1. Introduction; 2. Materials; 2.1. Cell Culture; 3. Method; 3.1. Troubleshooting; 3.2. Notes; References
  table of contents: Chapter 4: Measurement of intracellular Ca2+ mobilization to study GPCR signal transduction1. Introduction; 2. Materials; 2.1. Maintenance of Cell Lines; 2.2. Generation of "Stable Cell Lines"; 2.3. Fura2AM Loading and Spectrophotometric Analysis; 3. Methods; 3.1. Maintenance of Cell Lines; 3.2. Generation of "Stable Cell Lines"; 3.3. Fura2AM Loading; 3.4. Spectrophotometric Analysis; 4. Notes; References; Chapter 5: Visualization and quantification of GPCR trafficking in mammalian cells by confocal microscopy; 1. Introduction; 2. Methods; 2.1. Required Materials
  note: edited by Arun K. Shukla.
  note: Text in English.
  note: Includes bibliographical references and index.
  subject:
    topic: Cytology
    topic: Technique
  subject:
    topic: Cell receptors
  subject:
    topic: SCIENCE / Life Sciences / Anatomy & Physiology
  subject:
    topic: Cell receptors
  subject:
    topic: Cytology
    topic: Technique
  subject:
    topic: GTP-Binding Proteins
  subject:
    topic: Cytological Techniques
  subject:
    topic: Receptors, Cell Surface
  classification: QP552.G16 G67 2017eb
  classification: 571.6
  related item:
    title info:
      title: G protein-coupled receptors
    origin info:
      publisher: Cambridge, MA, USA : Academic Press is an imprint of Elsevier, 2017
      edition: Second edition.
  related item:
    title info:
      title: Methods in cell biology ; v. 142
  identifier: 9780128133200
  identifier: 0128133201
  identifier: 9780128133194
  identifier: 0128133198
  identifier: 1038723 MIL
  identifier: http://proxy.library.tamu.edu/login
  record info:
    record content source: N$T
    record creation date: 171004
    record change date: 20230131194853.0
    record identifier: in00004055196
    language of cataloging:
      language term: eng