Backbone resonance assignments of an artificially engineered TEM-1/PSE-4 Class A β-lactamase chimera.


Journal Article

The rapid evolution of Class A β-lactamases, which procure resistance to an increasingly broad panel of β-lactam antibiotics, underscores the urgency to better understand the relation between their sequence variation and their structural and functional features. To date, more than 300 clinically-relevant β-lactamase variants have been reported, and this number continues to increase. With the aim of obtaining insights into the evolutionary potential of β-lactamases, an artificially engineered, catalytically active chimera of the Class A TEM-1 and PSE-4 β-lactamases is under study by kinetics and NMR. Here we report the (1)H, (13)C and (15)N backbone resonance assignments for the 30 kDa chimera cTEM-17m. Despite its high molecular weight, the data provide evidence that this artificially-evolved chimeric enzyme is well folded. The hydrolytic activity of cTEM-17m was determined using the chromogenic substrate CENTA, with K (M) = 160 ± 35 μM and k (cat) = 20 ± 4 s(-1), which is in the same range as the values for TEM-1 and PSE-4 β-lactamases.

Full Text

Duke Authors

Cited Authors

  • Morin, S; Clouthier, CM; Gobeil, S; Pelletier, JN; Gagné, SM

Published Date

  • October 2010

Published In

Volume / Issue

  • 4 / 2

Start / End Page

  • 127 - 130

PubMed ID

  • 20383614

Pubmed Central ID

  • 20383614

Electronic International Standard Serial Number (EISSN)

  • 1874-270X

International Standard Serial Number (ISSN)

  • 1874-2718

Digital Object Identifier (DOI)

  • 10.1007/s12104-010-9227-8


  • eng