SPINE stands for strep–protein interaction experiment. SPINE is a powerful tool to detect
protein–protein interaction
Protein–protein interactions (PPIs) are physical contacts of high specificity established between two or more protein molecules as a result of biochemical events steered by interactions that include electrostatic forces, hydrogen bonding and th ...
s in vivo. The bait protein has to be expressed with a
Strep-tag
The Strep-tag system is a method which allows the purification and detection of proteins by affinity chromatography. The Strep-tag II is a synthetic peptide consisting of eight amino acids (Tryptophan, Trp-Serine, Ser-Histidine, His-Proline, Pro-G ...
under the conditions when the potential interaction partners are presumably present in the cells. The addition of
formaldehyde
Formaldehyde ( , ) (systematic name methanal) is a naturally occurring organic compound with the formula and structure . The pure compound is a pungent, colourless gas that polymerises spontaneously into paraformaldehyde (refer to section F ...
links the bait protein to its potential interaction partners. The bait protein together with its potential interaction partners can then be isolated using a Strep-Tactin Sepharose column. The cross-links between the bait protein and the potential interaction partner can be cleaved by heating the samples in Laemmli buffer. Finally, the co-purified interaction partner can be separated by SDS PAGE and identified by mass spectrometry.
References
* Herzberg C, Weidinger LA, Dörrbecker B, Hübner S, Stülke J and Commichau FM (2007)
SPINE: A method for the rapid detection and analysis of protein–protein interactions in vivo Proteomics 7(22):4032-4035
* Müller VS, Jungblut PR, Meyer TF, Hunke S (2011)
Membrane-SPINE: an improved method to identify protein–protein interaction partners of membrane proteins in vivo Proteomics 11(10):2124-2128
* Müller VS, Tschauner K, Hunke S (2013)
Membrane-SPINE: a biochemical tool to identify protein–protein interactions of membrane proteins in vivo J Vis Exp 7(81):e50810
Protein–protein interaction assays
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