KRAS, 1st ed. 2024 Methods and Protocols Methods in Molecular Biology Series, Vol. 2797
Coordonnateurs : Stephen Andrew G., Esposito Dominic
This volume details protocols ranging from high yield production metabolically labeled KRAS for NMR studies to approaches that quantify engagement of novel molecules that bind KRAS in live cells. Chapters focus on protein production and characterization, biochemical assays, cell-based assays, KRAS-membrane interactions, targeting KRAS, and cell models. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, KRAS: Methods and Protocols aims to provide methods that will be instrumental in the development of future clinically approved KRAS therapeutics.
1 KRAS as a therapeutic target
Dwight V. Nissley, Andrew G. Stephen, Ming Yi, and Frank McCormick
2 The abundance of KRAS and RAS gene mutations in cancer
Edward C. Stites
3 Production of isotopically labeled KRAS4b
Troy Taylor and William Gillette
4 Nucleotide Exchange on RAS Proteins using Hydrolysable and Non-Hydrolysable Nucleotides
Timothy Waybright
5 Crystallographic studies of KRAS in Complex with Small Molecules and RAS-binding Proteins
Albert H. Chan and Dhirendra K. Simanshu
6 Considerations around Structure-Based Drug Discovery for KRAS using DOCK
Mayukh Chakrabarti, Y. Stanley Tan, and Trent Balius
7 Measurement of KRAS GTPase activity
Dana Rabara and Andrew G. Stephen
8 Affinity Measurement of Non-covalent Interactions of the covalent KRAS G12C GDP inhibitor MRTX849 to RAS Isoforms using Surface Plasmon Resonance
Patrick Alexander and Andrew G. Stephen
9 1D and 2D NMR for KRAS:ligand binding
Gabriel Cornilescu
10 Luminescence based techniques for KRAS thermal stability monitoring
Kari Kopra
11 MALDI-TOF mass spectrometry-based assay for measuring covalent target engagement of KRAS G12C inhibitors
Marcin Dyba, John-Paul Denson, and Anna E. Maciag
12 KRAS4b:RAF-1 Homogenous Time-Resolved Fluorescence Resonance Energy Transfer Assay for Drug Discovery
Erik K. Larsen, Maria Abreu-Blanco, Dana Rabara, and Andrew G. Stephen
13 Biophysical characterization of RAS-SOS Complexes by Native Mass Spectrometry
Sangho Yun, Elena Scott, and Arthur Laganowsky
14 Profiling complex RAS-effector interactions using NMR spectroscopy
Regina Strakhova and Matthew J. Smith
15 Probing RAS Function Using Monobody and NanoBiT Technologies
Michael Whaby, Rakesh Sathish Nair, and John O'Bryan
16 Analysis of the guanine nucleotide-bound state of KRAS by ion-pair reversed-phase high-performance liquid chromatography
Makoto Araki and Kenji Kontani
17 Real-time monitoring of RAS activity using in vitro and in-cell NMR spectroscopy
Qingci Zhao, Ichio Shimada, and Noritaka Nishida
18 Studying RAS Interactions in live cells with BRET
John Columbus and Tommy Turbyville
19 FLIM-FRET Protein-Protein Interaction Assay
Pedro Andrade Bonilla and Rebika Shrestha
20 High-throughput cell-based screening of small molecule KRAS signaling inhibitors using a homogeneous time resolved fluorescence (HTRF) assay
Brian P. Smith, Megan Rigby, Roger Ma, and Anna E. Maciag
21 A method to conditionally measure target engagement at intracellular RAS and RAF complexes.
James D. Vasta, Ani Michaud, J. Aaron Crapster, and Matthew B. Robers
22 FLAG-KRAS4B as a model system for KRAS4B proteoform and PTM evaluation by mass spectrometry
Robert A. D’Ippolito, Grace M. Scheidemantle, Brian Smith, Katie Powell, Scott Eury, Abigail Neish, Jennifer Mehalko, Lauren Beaumont, Nicole Fer, Vanessa Wall, William Burgan, Anna E. Maciag, Dominic Esposito, and Caroline J. DeHart
23 Creation of an isogenic H/N/KRAS-less mouse embryonic fibroblast cell line panel derived from a size sorted diploid clone
William Burgan
24 Quality Control of an Isogenic H/N/KRAS-less Mouse Embryonic Fibroblast Cell Line Panel
Bingfang Xu and Katie Powell
25 A Facile Method to Append a Bio-ID Tag to Endogenous Mutant Kras Alleles
Konstantin Budagyan, Alexa C. Cannon, and Jonathan Chernoff
Includes cutting-edge methods and protocols
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
Date de parution : 04-2024
Ouvrage de 350 p.
17.8x25.4 cm
Disponible chez l'éditeur (délai d'approvisionnement : 15 jours).
Prix indicatif 232,09 €
Ajouter au panier