TY  - JOUR
AU  - Atkinson, Sarah C.
AU  - Dogovski, Con
AU  - Wood, Kathleen
AU  - Griffin, Michael D. W.
AU  - Gorman, Michael A.
AU  - Hor, Lilian
AU  - Reboul, Cyril F.
AU  - Buckle, Ashley M.
AU  - Wuttke, Joachim
AU  - Parker, Michael W.
AU  - Dobson, Renwick C. J.
AU  - Perugini, Matthew A.
TI  - Substrate Locking Promotes Dimer-Dimer Docking of an Enzyme Antibiotic Target
JO  - Structure
VL  - 26
IS  - 7
SN  - 0969-2126
CY  - Cambridge, Mass.
PB  - Cell Press
M1  - FZJ-2018-07248
SP  - 948 - 959.e5
PY  - 2018
AB  - Protein dynamics manifested through structural flex-ibility play a central role in the function of biologicalmolecules. Here we explore the substrate-mediatedchange in protein flexibility of an antibiotic targetenzyme, Clostridium botulinum dihydrodipicolinatesynthase. We demonstrate that the substrate, pyru-vate, stabilizes the more active dimer-of-dimers ortetrameric form. Surprisingly, there is little differencebetween the crystal structures of apo and substrate-bound enzyme, suggesting protein dynamics may beimportant. Neutron and small-angle X-ray scatteringexperiments were used to probe substrate-induceddynamics on the sub-second timescale, but nosignificant changes were observed. We thereforedeveloped a simple technique, coined protein dy-namics-mass spectrometry (ProD-MS), which en-ables measurement of time-dependent alkylation ofcysteine residues. ProD-MS together with X-raycrystallography and analytical ultracentrifugationanalyses indicates that pyruvate locks the conforma-tion of the dimer that promotes docking to the moreactive tetrameric form, offering insight into ligand-mediated stabilization of multimeric enzymes.
LB  - PUB:(DE-HGF)16
C6  - pmid:29804823
UR  - <Go to ISI:>//WOS:000437809100005
DO  - DOI:10.1016/j.str.2018.04.014
UR  - https://juser.fz-juelich.de/record/858361
ER  -