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Cdc48A pulldown from an amino link column baited with SAMP1

UID: 11108

Author(s): Dantuluri, Swathi*, Maupin-Furlow, Julie * MSK affiliated

Description
H. volcanii Cdc48a, RecJ3/4 and RNase J associate with Ubl-coated beads. A) Strategy to isolate H. volcanii proteins that bind agarose beads charged with monomeric Ubl (vs. BSA) as bait. Cell lysate was from triplicate cultures of H. volcanii NH02-pJAM957. B) Left, Non-reducing SDS-PAGE of H. volcanii proteins that bound BSA- (control, lane 1) vs. Ubl- (lane 2) decorated beads in the presence of ATP. Gels were stained with SYPRO Ruby. Red bar, HMW(250kDa) and LMW(50kDa) (high and low molecular weight) regions of gel excised from lanes 1 and 2 for LC- MS/MS analysis. Right, Proteins identified at > 99.9% probability and < 0.1 % false discovery rate (FDR) in the Ubl (not BSA) samples. Similar trend was observed in independent experiment. ATP was required for this association. Theor. Mr, theoretical molecular mass based on genome sequence. Normalized total spectra of sample minus control is indicated.
Subject of Study
Subject(s)
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Accession #: PXD019897

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Associated Publications
Equipment Used
LTQ Orbitrap
Software Used
Mascot
Scaffold
Dataset Format(s)
RAW, MGF
Dataset Size
268 MB
Data Catalog Record Updated
2024-01-03