Fast atom bombardment mass spectral search for the amino terminus of genetically engineered alpha 1-antitrypsin

Protein structure determination of genetically engineered alpha 1-antitrypsin was carried out using the technique of 'fast atom bombardment (FAB) MAPPING'. CNBr, tryptic and chymotryptic FAB MAPS were produced. The anticipated amino terminal region of the molecule was not mapped at the expected mass, raising the possibility of post-translational modification. A specific experiment was designed to isolate and identify this region by FAB mass spectral screening of high-performance liquid chromatography separated peptides. A signal at m/z 1231 was observed which could not be assigned to any sequence in the molecule using the computer program. Following CNBr treatment, this signal disappeared completely, giving rise to a new signal at m/z 1058. The amino terminus was thus found to be extended by the presence of an N-acetyl methionine residue, and this discovery is the subject of the present paper; another modification within the sequence will be reported elsewhere. Combining the FAB MAPPING data, the overall structural confirmation achieved was 93% of the recombinant alpha 1-antitrypsin molecule.