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Analysis of human AKT PH domain using limited proteolysis (LiP) followed by LC-MS/MS

ABSTRACT: We identified that a lncRNA named Long Intergenic Noncoding RNA for Kinase Activation (LINK-A, also LINC01139) directly and specifically interacts with AKT and PIP3. The LINK-A-AKT and -PIP3 interactions facilitate AKT recruitment and consequent activation. we also identified the single nucleotides of LINK-A required for PIP3 and AKT binding respectively. We hypothesized that the formation of LINK-A-PIP3-AKT complex may cause potential conformational changes of AKT PH domain, which can be studied by limited proteolysis. We applied limited proteolysis (LiP) followed by liquid chromatography-mass spectrometry (LC-MS/MS) analysis to analyze the AKT PH domain in the absence or presence of PIP3 and/or LINK-A RNA.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: chunru lin

LAB HEAD: Liuqing Yang

PROVIDER: PXD005636 | Pride | 2017-02-10

REPOSITORIES: Pride

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