Defining the architecture of KPC-2 carbapenemase

The rise of multi-drugresistanceinbacterial pathogensis one ofthe grandchallenges facing medical science.A main concern is the speed of development of KPC carbapenemases resistance, thus putting at risk theefficacy of the most recently approved inhibitors, including carbapenems and avibactam.New strategies toovercome resistance are urgentlyrequired, which will ultimately be facilitated by a deeperunderstanding of the mechanisms that regulate KPC carbapenemase function.Usingenhancedsamplingcomputational methods together with site directed mutagenesis,we report theidentification of hydrophobic networks , whose integrity has been found to be essentialfor KPC-2 resistance. Present throughout the structure, they are responsible for structural integrityand allosteric signalling and when disrupted lead to a complete loss of the KPC-2 mediatedresistance phenotype, leading to the recovery of β-lactam sensitivity.