Crystalline alcohol dehydrogenases from the mesophilic bacterium Clostridium beijerinckii and the thermophilic bacterium Thermoanaerobium brockii: Preparation, characterization and molecular symmetry

Two tetrameric NADP(+)-dependent bacterial secondary alcohol dehydrogenases have been crystallized in the apo- and the holoenzyme forms. Crystals of the hole-enzyme from the mesophilic Clostridium beijerinckii (NCBAD) belong to space group P2(1)2(1)2(1) with unit-cell dimensions a = 90.5, b = 127.9, c = 151.4 Angstrom. Crystals of the ape-enzyme (CBAD) belong to the same space group with unit-cell dimensions a = 80.4, b = 102.3, c = 193.5 Angstrom. Crystals of the hole-enzyme from the thermophilic Thermoanaerobium brockii (NTBAD) belong to space group P6(1(5)) (a = b = 80.6, c = 400.7 Angstrom). Crystals of the ape-form of TBAD (point mutant G198D) belong to space group P2(1) with cell dimensions a = 123.0, b = 84.8, c = 160.4 Angstrom beta = 99.5 degrees. Crystals of CBAD, NCBAD and NTBAD contain one tetramer per asymmetric unit. They diffract to 2.0 Angstrom resolution at liquid nitrogen temperature. Crystals of TBAD(G198D) have two tetramers per asymmetric unit and diffract to 2.7 Angstrom at 276 K. Self-rotation analysis shows that both enzymes are tetramers of 222 symmetry.