Posttranslational modification of the 20S proteasomal proteins of the archaeon Haloferax volcanii.
20S proteasomes are large, multicatalytic proteases that play an important role in intracellular protein degradation. The barrel-like architecture of 20S proteasomes, formed by the stacking of four heptameric protein rings, is highly conserved from archaea to eukaryotes. The outer two rings are composed of alpha-type subunits, and the inner two rings are composed of beta-type subunits. The halophilic archaeon Haloferax volcanii synthesizes two different alpha-type proteins, alpha1 and alpha2, and one beta-type protein that assemble into at least two 20S proteasome subtypes. In this study, we demonstrate that all three of these 20S proteasomal proteins (alpha1, alpha2, and beta) are modified either post- or cotranslationally. Using electrospray ionization quadrupole time-of-flight mass spectrometry, a phosphorylation site of the beta subunit was identified at Ser129 of the deduced protein sequence. In addition, alpha1 and alpha2 contained N-terminal acetyl groups. These findings represent the first evidence of acetylation and phosphorylation of archaeal proteasomes and are one of the limited examples of post- and/or cotranslational modification of proteins in this unusual group of organisms.
Humbard
Matthew A
MA
Department of Microbiology and Cell Science, University of Florida, Gainesville, 32611-0700, USA.
Stevens
Stanley M
SM
Maupin-Furlow
Julie A
JA
D000107:Acetylation; D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000378:metabolism*; D017124:Conserved Sequence; D015180:Electrophoresis, Gel, Two-Dimensional; D019622:Haloferax volcanii / Q000378:metabolism*; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000378:metabolism*; D010766:Phosphorylation; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry / Q000378:metabolism*; D020033:Protein Isoforms / Q000032:analysis; D046188:Protein Modification, Translational; D011499:Protein Processing, Post-Translational; D016415:Sequence Alignment; D021241:Spectrometry, Mass, Electrospray Ionization
D019843:Archaeal Proteins; D007527:Isoenzymes; D020033:Protein Isoforms; D010447:Peptide Hydrolases; D046988:Proteasome Endopeptidase Complex