%0 Journal Article
%A Atkinson, Sarah C.
%A Dogovski, Con
%A Wood, Kathleen
%A Griffin, Michael D. W.
%A Gorman, Michael A.
%A Hor, Lilian
%A Reboul, Cyril F.
%A Buckle, Ashley M.
%A Wuttke, Joachim
%A Parker, Michael W.
%A Dobson, Renwick C. J.
%A Perugini, Matthew A.
%T Substrate Locking Promotes Dimer-Dimer Docking of an Enzyme Antibiotic Target
%J Structure
%V 26
%N 7
%@ 0969-2126
%C Cambridge, Mass.
%I Cell Press
%M FZJ-2018-07248
%P 948 - 959.e5
%D 2018
%X 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.
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:29804823
%U <Go to ISI:>//WOS:000437809100005
%R 10.1016/j.str.2018.04.014
%U https://juser.fz-juelich.de/record/858361