TY  - JOUR
AU  - Schulte, Marianne
AU  - Petrović, Dušan
AU  - Neudecker, Philipp
AU  - Hartmann, Rudolf
AU  - Pietruszka, Jörg
AU  - Willbold, Sabine
AU  - Willbold, Dieter
AU  - Panwalkar, Vineet
TI  - Conformational Sampling of the Intrinsically Disordered C-Terminal Tail of DERA Is Important for Enzyme Catalysis
JO  - ACS catalysis
VL  - 8
IS  - 5
SN  - 2155-5435
CY  - Washington, DC
PB  - ACS
M1  - FZJ-2018-02552
SP  - 3971 - 3984
PY  - 2018
AB  - 2-Deoxyribose-5-phosphate aldolase (DERA) catalyzes the reversible conversion of acetaldehyde and glyceraldehyde-3-phosphate into deoxyribose-5-phosphate. DERA is used as a biocatalyst for the synthesis of drugs such as statins and is a promising pharmaceutical target due to its involvement in nucleotide catabolism. Despite previous biochemical studies suggesting the catalytic importance of the C-terminal tyrosine residue found in several bacterial DERAs, the structural and functional basis of its participation in catalysis remains elusive because the electron density for the last eight to nine residues (i.e., the C-terminal tail) is absent in all available crystal structures. Using a combination of NMR spectroscopy and molecular dynamics simulations, we conclusively show that the rarely studied C-terminal tail of E. coli DERA (ecDERA) is intrinsically disordered and exists in equilibrium between open and catalytically relevant closed states, where the C-terminal tyrosine (Y259) enters the active site. Nuclear Overhauser effect distance restraints, obtained due to the presence of a substantial closed state population, were used to derive the solution-state structure of the ecDERA closed state. Real-time NMR hydrogen/deuterium exchange experiments reveal that Y259 is required for efficiency of the proton abstraction step of the catalytic reaction. Phosphate titration experiments show that, in addition to the phosphate-binding residues located near the active site, as observed in the available crystal structures, ecDERA contains previously unknown auxiliary phosphate-binding residues on the C-terminal tail which could facilitate in orienting Y259 in an optimal position for catalysis. Thus, we present significant insights into the structural and mechanistic importance of the ecDERA C-terminal tail and illustrate the role of conformational sampling in enzyme catalysis.
LB  - PUB:(DE-HGF)16
UR  - <Go to ISI:>//WOS:000431727300029
C6  - pmid:30101036
DO  - DOI:10.1021/acscatal.7b04408
UR  - https://juser.fz-juelich.de/record/845271
ER  -