Characterizing proteins and their interactions in cells and tissues using the in situ proximity ligation assay.
The activity of proteins is typically regulated by secondary modifications and by interactions with other partners, resulting in the formation of protein complexes whose functions depend on the participating proteins. Accordingly, it is of central importance to monitor the presence of interaction complexes as well as their localization, thus providing information about the types of cells where the proteins are located and in what sub-cellular compartment these interactions occur. Several methods for visualizing protein interactions in situ have been developed during the last decade. These methods in most cases involve genetic constructs, and they have been successfully used in assays of living cell maintained in tissue culture, but they cannot easily be implemented in studies of clinical specimens. For such samples, affinity reagents like antibodies can be used to target the interacting proteins. In this review we will describe the in situ proximity ligation assays (in situ PLA), a method that is suitable for visualizing protein interactions in both tissue sections and in vitro cell lines, and we discuss research tasks when this or other method may be selected.
Söderberg
Ola
O
Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala University, SE-75185 Uppsala, Sweden. ola.soderberg@genpat.uu.se
Leuchowius
Karl-Johan
KJ
Gullberg
Mats
M
Jarvius
Malin
M
Weibrecht
Irene
I
Larsson
Lars-Gunnar
LG
Landegren
Ulf
U
D000906:Antibodies / Q000378:metabolism; D001681:Biological Assay / Q000379:methods; D002478:Cells, Cultured; D011088:DNA Ligases; D015342:DNA Probes / Q000493:pharmacokinetics*; D019281:Dimerization; D005347:Fibroblasts / Q000378:metabolism; D005456:Fluorescent Dyes / Q000493:pharmacokinetics; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods; D009841:Oligonucleotides / Q000378:metabolism; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000378:metabolism*; D012107:Research Design; D012680:Sensitivity and Specificity; D020798:Two-Hybrid System Techniques
D000906:Antibodies; D015342:DNA Probes; D005456:Fluorescent Dyes; D009841:Oligonucleotides; D011506:Proteins; D011088:DNA Ligases