Interacting proteins of AMPK studied using TurboID proximity labeling technology

Area of Science:

• Cellular Biology
• Molecular Biology
• Biochemistry

Background:

• AMP-activated protein kinase (AMPK) regulates cellular energy metabolism and is implicated in diseases like diabetes and myocardial infarction.
• Understanding AMPK's interactions is crucial for developing targeted therapeutics.

Purpose of the Study:

• To identify novel proteins interacting with AMPK using proximity labeling.
• To investigate the biological functions of these interacting proteins, particularly DNAJA1.

Main Methods:

• TurboID proximity labeling in U251 cells stably expressing AMPK-TurboID.
• Biotin labeling, silver staining, mass spectrometry (MS), western blotting, co-immunoprecipitation (co-IP), and immunofluorescence (IF).

Main Results:

• Successfully established stable cell lines and identified several AMPK-interacting proteins.
• DNAJ heat shock protein family (Hsp40) member A1 (DNAJA1) was validated as a key interacting protein.
• AMPK and DNAJA1 were shown to interact and synergistically protect cells from apoptosis.

Conclusions:

• DNAJA1 is a novel AMPK-interacting protein.
• The AMPK-DNAJA1 interaction offers a protective mechanism against cellular apoptosis.
• This finding may provide a basis for novel therapeutic strategies for metabolic and cardiovascular diseases.