Fast atom bombardment mass spectrometry and chemical analysis in determinations of acyl-blocked protein structures

• Published in: FEBS letters 1990-08, Vol.269 (1), p.194-196
• Main Authors: Egestad, Börje, Estonius, Mats, Danielsson, Olle, Persson, Bengt, Cederlund, Ella, Kaiser, Rudolf, Holmquist, Barton, Vallee, Bert, Parés, Xavier, Jefferey, Jonathan, Jörnvall, Hans
• Format: Article
• Language: English
• Subjects: Acetylation; Acylation; Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Blocked peptide; C-terminal determination; Enzymes and enzyme inhibitors; Fast atom bombardment; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Humans; Mass Spectrometry; mass spectroscopy; Molecular Sequence Data; N-terminal acetylation; Oxidoreductases - analysis; Protein Processing, Post-Translational; Proteins - analysis
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(90)81152-E

Peptide generation and fast atom bombardment mass spectrometry in combination with conventional chemical analysis was used to identify the blocking group and establish the N-terminal structure of six different proteins at the nanomole level. In this manner, the first terminal structures of three non-mammalian alcohol dehydrogenases were determined, demonstrating the presence of N-terminal acetylation in these piscine, amphibian, and avian enzymes. Similarly, two different yeast glucose-6-phosphate dehydrogenases and a minor variant of a human alcohol dehydrogenase were found to be acetylated. The exact end location of C-terminal structures was also established. Together, the analyses permit the definition of terminal regions and blocking groups, thus facilitating the delineation of remaining structures.