Fast atom bombardment mass spectrometry and chemical analysis in determinations of acyl-blocked protein structures.
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.
Egestad
B
B
Department of Physiological Chemistry, Karolinska Institutet, Stockholm, Sweden.
Estonius
M
M
Danielsson
O
O
Persson
B
B
Cederlund
E
E
Kaiser
R
R
Holmquist
B
B
Vallee
B
B
Parés
X
X
Jefferey
J
J
D000107:Acetylation; D000215:Acylation; D000595:Amino Acid Sequence; D000818:Animals; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000032:analysis; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis*
D011506:Proteins; D010088:Oxidoreductases