ABSTRACT
We report the measurement of the time of flight of â¼17 GeV ν(µ) on the CNGS baseline (732 km) with the Large Volume Detector (LVD) at the Gran Sasso Laboratory. The CERN-SPS accelerator has been operated from May 10th to May 24th 2012, with a tightly bunched-beam structure to allow the velocity of neutrinos to be accurately measured on an event-by-event basis. LVD has detected 48 neutrino events, associated with the beam, with a high absolute time accuracy. These events allow us to establish the following limit on the difference between the neutrino speed and the light velocity: -3.8 × 10(-6) < (v(ν)-c)/c < 3.1 × 10(-6) (at 99% C.L.). This value is an order of magnitude lower than previous direct measurements.
ABSTRACT
The role of three histidine residues (His205, His296 and His303) and Asp259, important for the catalysis of NAD+-specific D-lactate dehydrogenase, was investigated using site-directed mutagenesis. None of these residues is presumed to be involved in coenzyme binding because Km for NADH remained essentially unchanged for all the mutant enzymes. Replacement of His205 with lysine resulted in a 125-fold reduction in kcat and a slight lowering of the Km value for pyruvate. D259N mutant showed a 56-fold reduction in kcat and a fivefold lowering of Km. The enzymatic activity profile shifted towards acidic pH by approximately 2 units. The H303K mutation produced no significant change in kcat values, although Km for pyruvate increased fourfold. Substitution of His296 with lysine produced no significant change in kcat values or in Km for substrate. The results obtained suggest that His205 and Asp259 play an important role in catalysis, whereas His303 does not. This corroborates structural information available for some members of the D-specific dehydrogenases family. The catalytic His296, proposed from structural studies to be the active site acid/base catalyst, is not invariant. Its function can be accomplished by lysine and this has significant implications for the enzymatic mechanism.