SPINE: a method for the rapid detection and analysis of protein-protein interactions in vivo.
The detection and analysis of protein-protein interactions is one of the central tasks of proteomics in the postgenomic era. For this purpose, we present a procedure, the Strep-protein interaction experiment (SPINE) that combines the advantages of the Strep-tag protein purification system with those of reversible in vivo protein crosslinking by formaldehyde. Using two Bacillus subtilis regulator proteins, we demonstrate that this method is well suited to isolate protein complexes with high purity and virtually no background. Plasmids allowing the high-level expression of proteins carrying an N- or C-terminal Strep-tag in B. subtilis were constructed.
Herzberg
Christina
C
Department of General Microbiology, Georg-August-University Göttingen, Göttingen, Germany.
Weidinger
Lope Andrés Flórez
LA
Dörrbecker
Bastian
B
Hübner
Sebastian
S
Stülke
Jörg
J
Commichau
Fabian M
FM
D001412:Bacillus subtilis / Q000737:chemistry / Q000235:genetics; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D005557:Formaldehyde / Q000737:chemistry; D011485:Protein Binding; D040901:Proteomics / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D001426:Bacterial Proteins; D005557:Formaldehyde