SPINE (molecular biology)
Encyclopedia
SPINE stands for Strep-protein interaction experiment. SPINE is a powerful tool to detect protein-protein interaction
s in vivo. The bait protein has to be expressed with a Strep-tag
under the conditions when the potential interaction partners are presumably present in the cells. The addition of formaldehyde
links the bait protein to its potential interaction partners. The bait protein together with its potential interaction partners can then be isolated using a Streptactin 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.
Protein-protein interaction
Protein–protein interactions occur when two or more proteins bind together, often to carry out their biological function. Many of the most important molecular processes in the cell such as DNA replication are carried out by large molecular machines that are built from a large number of protein...
s in vivo. The bait protein has to be expressed with a Strep-tag
Strep-tag
The Strep-tag® system is a method which allows the purification and detection of proteins by affinity chromatography. The Strep-tag is a synthetic peptide consisting of eight amino acids...
under the conditions when the potential interaction partners are presumably present in the cells. The addition of formaldehyde
Formaldehyde
Formaldehyde is an organic compound with the formula CH2O. It is the simplest aldehyde, hence its systematic name methanal.Formaldehyde is a colorless gas with a characteristic pungent odor. It is an important precursor to many other chemical compounds, especially for polymers...
links the bait protein to its potential interaction partners. The bait protein together with its potential interaction partners can then be isolated using a Streptactin 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.