Cesium ion liquid secondary ion mass spectrometry of membrane-bound glycoproteins: Structural and topological considerations of acetylcholine receptor fromTorpedo californica

Abstract

We report mass mapping of a large (270 kD) multisubunit membrane bound glycoprotein, nicotinic acetylcholine receptor from Torpedo californica, using enzymic digests of the affinity purified whole receptor and cesium ion liquid secondary ion mass spectrometry. Peptides, glycopeptides and derivatized N-linked oligosaccharides were isolated by HPLC and identified by LSIMS. We have shown that mass spectrometric sensitivity is improved a hundred-fold through use of computer-controlled mass window stepping of an electro-optical multichannel array detection system on a LSIMS double focusing mass spectrometer. This new method permitted determination of the complete fragmentation pattern of Man8N2-ABEE using only 5 picomoles of sample.