AMP-activated protein kinase (AMPK) is a αβγ heterotrimer that plays a key role in regulating cellular energy metabolism. In particular it is known that binding of some small molecules to the β subunit increases enzyme activity. The γ subunit contains binding sites for adenine nucleotides which upon binding increase the activity of AMPK alleviating metabolic problems associated with several diseases. AMPK represents a promising drug target for type 2 diabetes, metabolic syndrome or obesity. Therefore, a detailed molecular understanding of the mechanism of small-molecule activation and regions that are involved in controlling the enzyme, are required. We are using two major analytical techniques to monitor protein dynamics: Nuclear Magnetic Resonance (NMR) spectroscopy and Mass Spectrometry (MS). NMR is being used to detect motions on the nanosecond to millisecond timescale with a focus on protein dynamics whereas MS is being used to collect data on long timescale motion. Since AMPK is a large protein and therefore difficult to analyze by NMR, we are mainly studying an 80 kDa and a 60kDa truncation of AMPK, both comprising the whole γ subunit and parts of the α and β subunits. The 80 kDa construct has already been investigated by NMR but it didn’t produce the expected resolution of the NMR spectrum. For attaining the 60 kDa truncation we used single site mutation for inserting a protease recognition sequence to cut the carbohydrate binding module (CBM) out of the enzyme. Furthermore some selected mutations of specific residues, known to be important for enzyme activity, will be tested. These constructs could be providing higher resolved outcomes of the enzyme dynamics by NMR and a deeper comprehension of the characteristic functionality of AMPK.