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

Backbone resonance assignments of an artificially engineered TEM-1/PSE-4 Class A β-lactamase chimera 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 1H, 13C and 15N 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Biomolecular NMR Assignments Springer Journals

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

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References (27)

Publisher
Springer Journals
Copyright
Copyright © 2010 by Springer Science+Business Media B.V.
Subject
Physics; Biochemistry, general; Polymer Sciences ; Biophysics and Biological Physics
ISSN
1874-2718
eISSN
1874-270X
DOI
10.1007/s12104-010-9227-8
pmid
20383614
Publisher site
See Article on Publisher Site

Abstract

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 1H, 13C and 15N 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.

Journal

Biomolecular NMR Assignments – Springer Journals

Published: Apr 10, 2010

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