2V20

Structure of a TEM-1 beta-lactamase insertant allosterically regulated by kanamycin and anions. Complex with sulfate.

Summary for 2V20

• Entry DOI: 10.2210/pdb2v20/pdb
• Related: 1AXB 1BT5 1BTL 1CK3 1ERM 1ERO 1ERQ 1ESU 1FQG 1JTD 1JTG 1JVJ 1JWP 1JWV 1JWZ 1LHY 1LI0 1LI9 1M40 1NXY 1NY0 1NYM 1NYY 1PZO 1PZP 1S0W 1TEM 1XPB 1XXM 1YT4 1ZG4 1ZG6 2V1Z
• Descriptor: BETA-LACTAMASE TEM, ZINC ION, SULFATE ION, ... (4 entities in total)
• Functional Keywords: hydrolase, insertion mutant, antibiotic resistance, allosteric regulation
• Biological source: ESCHERICHIA COLI
• Total number of polymer chains: 1
• Total formula weight: 32704.58

Authors

Evrard, C.,Barrios, H.,Mathonet, P.,Soumillion, P.,Fastrez, J.,Declercq, J.P. (deposition date: 2007-05-31, release date: 2008-06-24, Last modification date: 2024-11-20)

Primary citation

Volkov, A.N.,Barrios, H.,Mathonet, P.,Evrard, C.,Ubbink, M.,Declercq, J.P.,Soumillion, P.,Fastrez, J.
Engineering an Allosteric Binding Site for Aminoglycosides Into Tem1-Beta-Lactamase.
Chembiochem, 12:904-, 2011

PubMed Abstract: Allosteric regulation of enzyme activity is a remarkable property of many biological catalysts. Up till now, engineering an allosteric regulation into native, unregulated enzymes has been achieved by the creation of hybrid proteins in which a natural receptor, whose conformation is controlled by ligand binding, is inserted into an enzyme structure. Here, we describe a monomeric enzyme, TEM1-β-lactamase, that features an allosteric aminoglycoside binding site created de novo by directed-evolution methods. β-Lactamases are highly efficient enzymes involved in the resistance of bacteria against β-lactam antibiotics, such as penicillin. Aminoglycosides constitute another class of antibiotics that prevent bacterial protein synthesis, and are neither substrates nor ligands of the native β-lactamases. Here we show that the engineered enzyme is regulated by the binding of kanamycin and other aminoglycosides. Kinetic and structural analyses indicate that the activation mechanism involves expulsion of an inhibitor that binds to an additional, fortuitous site on the engineered protein. These analyses also led to the defining of conditions that allowed an aminoglycoside to be detected at low concentration.

PubMed: 21425229
DOI: 10.1002/CBIC.201000568

Experimental method

X-RAY DIFFRACTION (1.67 Å)