LaBaer Lab | Technology
Coupling NAPPA and Mass Spectrometry (MS) to enable many vs. many screens of protein interaction
Investigators: Sanjeeva Srivasta, Ph.D., Manuel Fuentes, Ph.D.
Collaborator: Sarah Volk and Philip Andrews (University of Michigan)
In this project we are coupling NAPPA protein array technique with mass spectrometry. A modified NAPPA chemistry, which is used on gold for surface plasmon resonance is used for this application. A test array is printed on gold slides along with K-coil capture peptide and proteins are expressed by using cell free expression system. Digestion and washing steps are performed prior to analysis using MALDI-TOF and TOF-TOF instruments. By using this approach we are able to detect MS and MS-MS spectra (Figure 1). In addition to generating MS-MS spectra, once this technology platform is established it can be used for several applications, including identification of peptide sequences for potential phosphorylation as well as an extremely high throughput method for detection of protein-protein interactions.
Coupling NAPPA and Mass Spectrometry. The modified NAPPA chemistry based on E-coil/K-coil is used to print spots on gold slides, and proteins are expressed by using cell free expression system. Proteins are trypsinzed and analyzed using MALDI-TOF and TOF-TOF instruments. The MS and MS-MS spectra are detected by using this approach.