Protein-Protein Interactions (2^nd Ed., Softcover reprint of the original 2nd ed. 2015) Methods and Applications Methods in Molecular Biology Series, Vol. 1278

The second edition covers a wide range of protein-protein interaction detection topics. Protein-Protein Interactions: Methods and Applications focuses on core technological platforms used to study protein-protein interactions and cutting-edge technologies that reflect recent scientific advances and the emerging focus on therapeutic discovery. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of necessary materials and reagents, step-by-step laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. These well-detailed protocols describe methods for identifying protein-protein interaction partners, analyzing of protein-protein interactions quantitatively and qualitatively, monitoring protein-protein interactions in live cells, and predicting and determining interaction interfaces.

Authoritative and cutting-edge, Protein-Protein Interactions: Methods and Applications, Second Edition is a valuable resource that will enable readers to elucidate the mechanisms of protein-protein interactions, determine the role of these interactions in diverse biological processes, and target protein-protein interactions for therapeutic discovery.

PART I    OVERVIEWS                    

1              Structural basis of protein-protein interactions

Robert Liddington

2              Quantitative analysis of protein-protein interactions        

Ziad Eletr and Keith Wilkinson

3              Protein-protein interaction databases      

Damian Szklarczyk and Lars Juhl Jensen

4              Computational prediction of protein-protein interactions 

Tobias Ehrenberger, Lewis C. Cantley, and Michael B. Yaffe

5              Structure-based computational approaches for small-molecule modulation of protein-protein interactions

David Xu, Bo Wang, and Samy O. Meroueh

6              Targeting protein-protein interactions for drug discovery            

David C. Fry

PART II   LABEL-FREE BIOSENSORS AND TECHNIQUES                    

7              Studying protein-protein interactions using surface plasmon resonance

Zenata Nikolovska-Coleska

8              Resonant waveguide grating for monitoring bimolecular interactions    

Meng Wu and Min Li

9              Quartz microbalance technology for probing biomolecular interactions 

Gabriella T. Heller, Alison R. Mercer-Smith, and Malkiat S. Johal

10           Label-free kinetic analysis of an antibody-antigen interaction using biolayer interferometry

Sriram Kumaraswamy and Renee Tobias

11           Characterization of protein-protein interactions by isothermal titration calorimetry

Adrian Velazquez-Campoy, Stephanie A. Leavitt, and Ernesto Freiere

12           Sedimentation equilibrium studies           

Ian A. Taylor, John F. Eccleston, and Katrin Rittinger

13           Detecting protein-protein interactions by gel filtration chromatography

Yan Bai

14           Using light scattering to determine the stoichiometry of protein complexes       

Jeremy Mogridge

15           Circular dichroism (CD) analyses of protein-protein interactions           

Norma Greenfield

16           Protein-protein interaction analysis by nuclear magnetic resonance spectroscopy

Peter M. Thompson, Moriah R. Beck, and Sharon L. Campbell

17           Quantitative protein analysis by mass spectrometry      

Vishwajeeth Pagala, Anthony High, Xusheng Wang, Haiyan Tan, Kiran Kodali, Ashutosh Mishra, Kanisha Kavdia, Yanji Xu, Zhiping Wu, and Junmin Peng

18           Using peptide arrays created by the SPOT method for defining protein-protein interactions

Yun Young Yim, Katherine Betke, and Heidi Hamm

PART III  TAG/AFFINITY-BASED METHODS                    

19           Fluorescence polarization assay to quantify protein-protein interactions            

Ronald T. Raines

20           Förster resonance energy transfer (FRET) microscopy for monitoring biomolecular interactions        

Alexa L. Mattheyses and Adam I. Marcus

Zusen Weng and Qinjian Zhao

22           Glutathione-S-transferase (GST)-fusion based assays for studying protein-protein interactions          

Haris G. Vikis and Kun-Liang Guan

23           Hexahistidine (6xHis) fusion-based assays for protein-protein interactions       

Mary C. Puckett

24           Studying protein-protein interactions via blot overlay/Far Western blot

Randy A. Hall

25           Co-immunoprecipitation from transfected cells  

Yoshinori Takahashi

26           In vivo protein cross-linking         

Fabrice Agou and Michel Véron

PART IV   CELL-BASED BIMOLECULAR INTERACTION REPORTER ASSAYS                      

27           Identification of protein-protein interactions by standard Gal4p-based yeast two-hybrid screening

Jeroen Wagemans and Rob Lavigne

28           Reverse two-hybrid techniques in the yeast Saccharomyces cerevisiae          

Matthew A. Bennett, Jack F. Shern, and Richard A. Kahn

29           MAPPIT, a mammalian two-hybrid method for in-cell detection of protein-protein interactions

Irma Lemmens, Sam Lievens, and Jan Tavernier

30           Bioluminescence resonance energy transfer to detect protein-protein interactions in live cells

Nicole E. Brown, Joe B. Blumer, and John R. Hepler

31           Mapping biochemical networks with protein fragment complementation assays          

Ingrid Remy and Stephen W. Michnick

32           Detection of protein-protein interaction using bimolecular fluorescence complementation assay

Cau D. Pham

33           Split-luciferase complementation assay to detect channel-protein interactions in live cells       

Alexander S. Shavkunov, Syed R. Ali, Neli I. Panova-Elektronova, and Fernanda Laezza

34           Confocal microscopy for intracellular co-localization of proteins           

Toshiyuki Miyashita

PART V  HIGH THROUGHPUT SCREENING ASSAYS FOR PROTEIN-  PROTEIN INTERACTIONS - CASE STUDIES                   

35           Fluorescence polarization assay to quantify protein-protein interactions in an HTS format

36           Estrogen receptor alpha/co-activator interaction assay - TR-FRET       

Terry W. Moore, Jillian R. Gunther, and John A. Katzenellenbogen

37           High content screening biosensor assay to identify disruptors of p53-hDM2 protein-protein interactions

Yun Hua, Christopher J. Strock. and Paul A. Johnston

38           Discovery of inhibitors of the MDM2-p53 protein-protein interaction

Liu Liu, Denzil Bernard, and Shaomeng Wang

39           Biophysical methods for identifying fragment-based inhibitors of protein-protein interactions

Samuel J. Pfaff, Michael S. Chimenti, Mark J. S. Kelly, and Michelle R. Arkin

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts