TY - JOUR
T1 - Nuclear proteasomes carry a constitutive posttranslational modification which derails SDS-PAGE (but not CTAB-PAGE)
AU - Pitcher, David S.
AU - De Mattos-Shipley, Kate M J
AU - Wang, Ziming
AU - Tzortzis, Konstantinos
AU - Goudevenoua, Katerina
AU - Flynn, Helen
AU - Bohn, George
AU - Rahemtulla, Amin
AU - Snijders, Ambrosius P.
AU - Karadimitris, Anastasios
AU - Kleijnen, Maurits F.
PY - 2014/9/1
Y1 - 2014/9/1
N2 - We report that subunits of human nuclear proteasomes carry a previously unrecognised, constitutive posttranslational modification. Subunits with this modification are not visualised by SDS-PAGE, which is used in almost all denaturing protein gel electrophoresis. In contrast, CTAB-PAGE readily visualises such modified subunits. Thus, under most experimental conditions, with identical samples, SDS-PAGE yielded gel electrophoresis patterns for subunits of nuclear proteasomes which were misleading and strikingly different from those obtained with CTAB-PAGE. Initial analysis indicates a novel modification of a high negative charge with some similarity to polyADP-ribose, possibly explaining compatibility with (positively-charged) CTAB-PAGE but not (negatively-charged) SDS-PAGE and providing a mechanism for how nuclear proteasomes may interact with chromatin, DNA and other nuclear components.
AB - We report that subunits of human nuclear proteasomes carry a previously unrecognised, constitutive posttranslational modification. Subunits with this modification are not visualised by SDS-PAGE, which is used in almost all denaturing protein gel electrophoresis. In contrast, CTAB-PAGE readily visualises such modified subunits. Thus, under most experimental conditions, with identical samples, SDS-PAGE yielded gel electrophoresis patterns for subunits of nuclear proteasomes which were misleading and strikingly different from those obtained with CTAB-PAGE. Initial analysis indicates a novel modification of a high negative charge with some similarity to polyADP-ribose, possibly explaining compatibility with (positively-charged) CTAB-PAGE but not (negatively-charged) SDS-PAGE and providing a mechanism for how nuclear proteasomes may interact with chromatin, DNA and other nuclear components.
M3 - Article (Academic Journal)
SN - 0006-3002
VL - 1844
SP - 2222
JO - Biochimica et Biophysica Acta (BBA)
JF - Biochimica et Biophysica Acta (BBA)
IS - 12
ER -