Proteomics

Dataset Information

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AKT2 interacting proteins_ MBP- and MAP-TAPs


ABSTRACT: Tagged AKT2 was expressed in HEK293T cells. For quantification SILAC labeling was performed. MBP-TAP: Cell lysates were mixed at a 1:1 ratio with unlabeled wild type cells before Tandem Affinity Purification. MAP-TAP: Same amount of labeled and unlabeled cell lysates were purified via Tandem Affinity Purification and afterwards eluates were mixed at a 1:1 ratio. AKT2 and co-purified proteins were digested with trypsin, fractionated via SCX and analyzed via LC-MS. Sequence database-search of the MS data was performed against the uniprot human taxonomy-9606 database containing 83659 entries using the SEQUEST algorithm with the following parameters: trypsin specificity, two missed cleavage sites, precurser ion mass accuracy tolerance of 10–30 ppm, cysteine carbamidomethylation, methionine oxidation, pSTY, N-terminal protein acetylation and, when performed, SILAC labels Lys-6, Arg-6 specified as modifications. The minimal cross-correlation score (XCorr) was set to 2.0, 2.5 and 3.0 for charge states +2, +3 and +4 respectively. The Delta Cn had to be >0.1 and the minimal peptide probability allowed was 0.05. The minimum number of peptides necessary for protein identification was three.

INSTRUMENT(S): LTQ Orbitrap, instrument model

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hek-293 Cell, Hek-293t Cell

SUBMITTER: Katharina Bottermann  

PROVIDER: PXD000197 | Pride | 2013-08-13

REPOSITORIES: Pride

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Publications

Systematic Analysis Reveals Elongation Factor 2 and α-Enolase as Novel Interaction Partners of AKT2.

Bottermann Katharina K   Reinartz Michael M   Barsoum Marian M   Kötter Sebastian S   Gödecke Axel A  

PloS one 20130618 6


AKT2 is one of the three isoforms of the protein kinase AKT being involved in the modulation of cellular metabolism. Since protein-protein interactions are one possibility to convey specificity in signal transduction, we performed AKT2-protein interaction analysis to elucidate their relevance for AKT2-dependent cellular functions. We identified heat shock protein 90 kDa (HSP90), Cdc37, heat shock protein 70 kDa (HSP70), 78 kDa glucose regulated protein (GRP78), tubulin, GAPDH, α-enolase and elon  ...[more]

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