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Dual multimodular class A penicillin-binding proteins in Mycobacterium leprae
S Lepage1, P Dubois, T K Ghosh
1Centre d'Ingénierie des Protéines, Institut de Chimie, Université dede Liège, Belgium.
Journal of Bacteriology
|July 1, 1997
Summary
Mycobacterium leprae penicillin-binding protein 1 (PBP1) was produced and purified. This class A PBP1 shows high affinity for penicillin and catalyzes acyl transfer reactions, unlike other class A PBPs.
Area of Science:
- Microbiology
- Molecular Biology
- Biochemistry
Background:
- The Mycobacterium leprae genome library contains the ponA gene.
- This gene encodes a multimodular class A penicillin-binding protein (PBP), designated PBP1.
Purpose of the Study:
- To produce and purify M. leprae PBP1.
- To characterize the biochemical properties of M. leprae PBP1, including its catalytic activity and penicillin-binding affinity.
Main Methods:
- The ponA gene was expressed in Escherichia coli.
- Recombinant PBP1 was extracted from membranes using CHAPS detergent.
- Purification was achieved via Ni2(+)-nitrilotriacetic acid-agarose chromatography.
Main Results:
- Purified M. leprae PBP1 was obtained.
- PBP1 catalyzes acyl transfer reactions on thiolesters.
- PBP1 exhibits high affinity for penicillin.
- M. leprae PBP1 denatures above 25°C, unlike other class A PBPs.
Conclusions:
- M. leprae PBP1 is a functional class A penicillin-binding protein.
- Its unique properties, including thermal instability and high penicillin affinity, warrant further investigation for potential therapeutic targets.