SAN ANTONIO, June 8 (GenomeWeb News) - A University of Kentucky researcher has come up with a high-throughput protocol for preparing membrane proteins for mass spectrometry analysis using only the common reagents associated with regular, in-gel protein digestion.
Membrane proteins are traditionally hard to analyze because they contain hydrophobic portions that can only be solubulized using detergents or organic solvents.
The trick to the new method, called the "Tube-Gel" digestion protocol, is that hydrophobic proteins dissolved in detergent are mixed with acrylamide solution before it is polymerized, and then poured into a thin tube, said Haining Zhu, the assistant professor at the
GenomeWeb News spoke with Zhu at the American Society for Mass Spectrometry's 2005 meeting, held here this week.
When compared to the traditional membrane protein protocol, which involves pouring an acrylamide slab gel, loading the solubulized proteins into lanes on the gel, running the gel, washing the gel, then slicing the gel into pieces before protein digestion inside the gel pieces, the new Tube-Gel method is much more high throughput, Zhu noted. However, the new method did result in about 100 less proteins being identified by mass spec, when compared side-by-side with the traditional membrane protein protocol.
"You can go home and use [the protocol]. It does not cost anything extra," said Zhu.
Zhu said he had considered patenting the new method, but it would be difficult to patent something that is so readily accessible.
A paper describing Zhu's new method is under review by the journal Molecular and Cellular Proteomics.
Tien Shun Lee is the editor of ProteoMonitor, a GenomeWeb News publication.