Nuclear membrane proteins with potential disease links found by subtractive proteomics

To comprehensively identify integral membrane proteins of the nuclear envelope (NE), we prepared separatety NEs and organdies known to cofractionate with them from liver. Proteins detected by multidimensional protein identification technology in the cofractionating organelles were subtracted from the NE data set. In addition to all 13 known NE integral proteins, 67 uncharacterized open reading frames with predicted membrane-spanning regions were identified. All of the eight proteins tested targeted to the NE, indicating that there are substantially more integral proteins of the NE than previously thought. Furthermore, 23 of these mapped within chromosome regions linked to a variety of dystrophies.
To identify integral proteins of the NE, we took advantage of recent advances in high-throughput shotgun proteomics using multidimensional protein identification technology (MudPIT) (4,5), by which the coupling of tandem [...]