Enzyme dynamics during catalysis measured by NMR spectroscopy.
Many biological processes, in particular enzyme catalysis, occur in the microsecond to millisecond time regime. While the chemical events and static structural features of enzyme catalysis have been extensively studied, very little is known about dynamic processes of the enzyme during the catalytic cycle. Dynamic NMR methods such as ZZ-exchange, line-shape analysis, Carr-Purcell-Meiboom-Gill (CPMG), and rotating frame spin-lattice relaxation (R(1rho)) experiments are powerful in detecting conformational rearrangements with interconversion rates between 0.1 and 10(5) s(-1). In this chapter, the first application of these methods to enzymes during catalysis is described, in addition to studies on several other enzymes in their free states and in complex with ligands. From the experimental results of all systems, a picture arises in which flexibility in the microsecond to millisecond time regime is intrinsic and likely to be an essential property of the enzyme. Quantitative analysis of dynamics at multiple sites of the enzyme reveal large-scale collective motions. For several enzymes, the frequency of motion is comparable to the overall turnover rate, raising the possibility that conformational rearrangements may be rate limiting for catalysis in these enzymes.
Kern
Dorothee
D
Department of Biochemistry, Brandeis University, Waltham, Massachusetts 02454, USA.
Eisenmesser
Elan Z
EZ
Wolf-Watz
Magnus
M
D000263:Adenylate Kinase / Q000737:chemistry / Q000378:metabolism; D036022:Capsid Proteins / Q000737:chemistry / Q000378:metabolism; D002384:Catalysis; D021984:Cyclophilin A / Q000737:chemistry / Q000378:metabolism; D004798:Enzymes / Q000737:chemistry* / Q000378:metabolism*; D015497:HIV-1 / Q000737:chemistry / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D009587:Nitrogen Isotopes; D017434:Protein Structure, Tertiary
D036022:Capsid Proteins; D004798:Enzymes; D009587:Nitrogen Isotopes; D000263:Adenylate Kinase; D021984:Cyclophilin A