1UR5

Stabilization of a Tetrameric Malate Dehydrogenase by Introduction of a Disulfide Bridge at the Dimer/Dimer Interface

Summary for 1UR5

• Entry DOI: 10.2210/pdb1ur5/pdb
• Related: 1GUY
• Descriptor: MALATE DEHYDROGENASE, NICOTINAMIDE-ADENINE-DINUCLEOTIDE, CADMIUM ION, ... (6 entities in total)
• Functional Keywords: oxidoreductase, tricarboxylic acid cycle, malate dehydrogenase
• Biological source: CHLOROFLEXUS AURANTIACUS
• Total number of polymer chains: 2
• Total formula weight: 67582.31

Authors

Bjork, A.,Dalhus, B.,Mantzilas, D.,Eijsink, V.G.H.,Sirevag, R. (deposition date: 2003-10-27, release date: 2003-11-05, Last modification date: 2024-10-16)

Primary citation

Bjork, A.,Dalhus, B.,Mantzilas, D.,Eijsink, V.G.H.,Sirevag, R.
Stabilization of a Tetrameric Malate Dehydrogenase by Introduction of a Disulfide Bridge at the Dimer-Dimer Interface
J.Mol.Biol., 334:811-, 2003

PubMed Abstract: Malate dehydrogenase (MDH) from the moderately thermophilic bacterium Chloroflexus aurantiacus (CaMDH) is a tetrameric enzyme, while MDHs from mesophilic organisms usually are dimers. To investigate the potential contribution of the extra dimer-dimer interface in CaMDH with respect to thermal stability, we have engineered an intersubunit disulfide bridge designed to strengthen dimer-dimer interactions. The resulting mutant (T187C, containing two 187-187 disulfide bridges in the tetramer) showed a 200-fold increase in half-life at 75 degrees C and an increase of 15 deg. C in apparent melting temperature compared to the wild-type. The crystal structure of the mutant (solved at 1.75 A resolution) was essentially identical with that of the wild-type, with the exception of the added inter-dimer disulfide bridge and the loss of an aromatic intra-dimer contact. Remarkably, the mutant and the wild-type had similar temperature optima and activities at their temperature optima, thus providing a clear case of uncoupling of thermal stability and thermoactivity. The results show that tetramerization may contribute to MDH stability to an extent that depends strongly on the number of stabilizing interactions in the dimer-dimer interface.

PubMed: 14636605
DOI: 10.1016/J.JMB.2003.10.006

Experimental method

X-RAY DIFFRACTION (1.75 Å)