Structural transformations in the dynamics of Michaelis complex formation in lactate dehydrogenase.
The dynamical nature of the binding of a substrate surrogate to lactate dehydrogenase is examined on the nanoseconds to milliseconds timescale by laser-induced temperature-jump relaxation spectroscopy. Fluorescence emission of the nicotinamide group of bound NADH is used to define the pathway and kinetics of substrate binding. Assignment of specific kinetic states and elucidation of their structures are accomplished using isotope edited infrared absorption spectroscopy. Such studies are poised to yield a detailed picture of the coupling of protein dynamics to function.
McClendon
Sebastian
S
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York, 10461, USA.
Vu
Dung M
DM
Clinch
Keith
K
Callender
Robert
R
Dyer
R Brian
RB
D001665:Binding Sites; D001703:Biophysics / Q000379:methods*; D002384:Catalysis; D006860:Hydrogen Bonding; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000737:chemistry*; D007834:Lasers; D008956:Models, Chemical; D009243:NAD / Q000737:chemistry; D009536:Niacinamide / Q000737:chemistry; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D017550:Spectroscopy, Fourier Transform Infrared; D013379:Substrate Specificity; D013696:Temperature; D013997:Time Factors
D011506:Proteins; D009243:NAD; D009536:Niacinamide; D007770:L-Lactate Dehydrogenase