Table 3.

Main experimental techniques used for reconstruction or validation of protein-protein interaction networks.

Technique	Large scale implementation	Binary interaction or complex	Advantage	Disadvantage	Organisms	References
Y2H - Yeast two hybrid	+++	B	No antibody required	Elevated rate of false-positives; Nuclear localization of proteins	Francisella tularensis; Blumeria Graminis; Pseudomonas syringae; Hyaloperonospora arabidopsidis; Golovinomyces orontii	Weßling et al., 2014; Wallqvist et al., 2015; Pennington et al., 2016
PCA - Protein-fragment complementation Assay	++	C	Interaction with membrane proteins	Works better with small monomeric proteins	Vibrio cholerae; Escherichia coli	Ozawa et al., 2001; Hatzios et al., 2012
FRET - Förster resonance energy transfer	+	B	Reversible interaction	Decreased sensibility; Photobleaching	Hordeum vulgare	Bhat et al., 2005
BiFC - Bimolecular fluorescence complementation	+++	B	Used for localization in living cells	Detection of weakly associated proteins	Agrobacterium tumefaciens	Lacroix et al., 2005
TAP - Tandem affinity purification-mass spectroscopy	+	C	Accurate and efficient for multiprotein complex	High experimental effort and extensive data analysis	Measles morbillivirus; Candida albicans	Kaneko et al., 2004; Komarova et al., 2011
Protein array	++	C	Highly specific recognition	Needs a set of labeled proteins	Staphylococcus Aureus	Scietti et al., 2016
Pull - down	+++	C	Medium level of standardization	Protein GST fusion may cause sterical hindrance	Streptococcus suis	Li et al., 2016
Phage display	+++	C	Great diversity of variant proteins that can be represented in a phage library	Post-translational modifications; selection condition of library	Helicobacter pylori	Jonsson et al., 2004