Electrospray mass spectrometry characterization of post-translational modifications of barley alpha-amylase 1 produced in yeast.
We have used electrospray mass spectrometry (ESMS) in combination with protein chemistry and genetics to delineate post-translational modifications in yeast of barley alpha-amylase 1 (AMY1), a 45 kD enzyme crucial for production of malt, an important starting material in the manufacture of beer and whisky. In addition to signal peptide processing these modifications are: (1) removal of C-terminal Arg-Ser by Kex1p, (2) glutathionylation of Cys95, (3) O-glycosylation, and (4) additional degradation of the C-terminus.
Søgaard
M
M
Department of Chemistry, Carlsberg Laboratory, Copenhagen Valby, Denmark.
Andersen
J S
JS
Roepstorff
P
P
Svensson
B
B
D001483:Base Sequence; D015153:Blotting, Western; D005978:Glutathione / Q000378:metabolism; D001467:Hordeum / Q000201:enzymology*; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D000516:alpha-Amylases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D011994:Recombinant Proteins; D000516:alpha-Amylases; D005978:Glutathione