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Methods for the detection and analysis of protein-protein interactions

Berggård, Tord LU ; Linse, Sara LU and James, Peter LU (2007) In Proteomics 7(16). p.2833-2842
Abstract
A large number of methods have been developed over the years to study protein-protein interactions. Many of these techniques are now available to the nonspecialist researcher thanks to new affordable instruments and/or resource centres. A typical protein-protein interaction study usually starts with an initial screen for novel binding partners. We start this review by describing three techniques that can be used for this purpose: (i) affinity-tagged proteins (ii) the two-hybrid system and (iii) some quantitative proteomic techniques that can be used in combination with, e.g., affinity chromatography and coimmunoprecipitation for screening of protein-protein interactions. We then describe some public protein-protein interaction databases... (More)
A large number of methods have been developed over the years to study protein-protein interactions. Many of these techniques are now available to the nonspecialist researcher thanks to new affordable instruments and/or resource centres. A typical protein-protein interaction study usually starts with an initial screen for novel binding partners. We start this review by describing three techniques that can be used for this purpose: (i) affinity-tagged proteins (ii) the two-hybrid system and (iii) some quantitative proteomic techniques that can be used in combination with, e.g., affinity chromatography and coimmunoprecipitation for screening of protein-protein interactions. We then describe some public protein-protein interaction databases that can be searched to identify previously reported interactions for a given bait protein. Four strategies for validation of protein-protein interactions are presented: confocal microscopy for intracellular colocalization of proteins, coimmunoprecipitation, surface plasmon resonance (SPR) and spectroscopic studies. Throughout the review we focus particularly on the advantages and limitations of each method. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
affinity tags, protein-protein interactions, purification, protein, protein complexes
in
Proteomics
volume
7
issue
16
pages
2833 - 2842
publisher
John Wiley & Sons
external identifiers
  • wos:000249295500012
  • scopus:34548418142
ISSN
1615-9861
DOI
10.1002/pmic.200700131
language
English
LU publication?
yes
id
213ba1b7-4fce-4f5b-aa3c-86882d5b3c2e (old id 687018)
date added to LUP
2007-12-04 16:29:40
date last changed
2017-11-19 03:39:34
@article{213ba1b7-4fce-4f5b-aa3c-86882d5b3c2e,
  abstract     = {A large number of methods have been developed over the years to study protein-protein interactions. Many of these techniques are now available to the nonspecialist researcher thanks to new affordable instruments and/or resource centres. A typical protein-protein interaction study usually starts with an initial screen for novel binding partners. We start this review by describing three techniques that can be used for this purpose: (i) affinity-tagged proteins (ii) the two-hybrid system and (iii) some quantitative proteomic techniques that can be used in combination with, e.g., affinity chromatography and coimmunoprecipitation for screening of protein-protein interactions. We then describe some public protein-protein interaction databases that can be searched to identify previously reported interactions for a given bait protein. Four strategies for validation of protein-protein interactions are presented: confocal microscopy for intracellular colocalization of proteins, coimmunoprecipitation, surface plasmon resonance (SPR) and spectroscopic studies. Throughout the review we focus particularly on the advantages and limitations of each method.},
  author       = {Berggård, Tord and Linse, Sara and James, Peter},
  issn         = {1615-9861},
  keyword      = {affinity tags,protein-protein interactions,purification,protein,protein complexes},
  language     = {eng},
  number       = {16},
  pages        = {2833--2842},
  publisher    = {John Wiley & Sons},
  series       = {Proteomics},
  title        = {Methods for the detection and analysis of protein-protein interactions},
  url          = {http://dx.doi.org/10.1002/pmic.200700131},
  volume       = {7},
  year         = {2007},
}