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submitted on 01.11.2019 and posted on 07.11.2019by Chun-Chi Chen, Satish Malwal, Xu Han, Weidong Liu, Lixin Ma, Chao Zhai, Longhai Dai, Jian-Wen Huang, Alli Shillo, Janish Desai, Xianqiang Ma, Yonghui Zhang, Rey-Ting Guo, Eric Oldfield
We report the structures of
the terpene cyclases Santalum album L. sesquisabinene synthases 1 and 2
and of santalene synthase, in apo forms, and with the sesquisabinene synthases,
bound to either farnesyl diphosphate (FPP), farnesyl S-thiolo-diphosphate, FPP
containing a POP bridging O-to-CCl2 substitution, or to sabinene,
leading to a sequential mechanism for substrate binding and catalysis. We
trapped early pre-catalytic inactive open forms that show how ligands initially
bind to the apo-proteins, then when the pocket closes, catalysis can proceed.
We also show that there are strong structural similarities between the most
highly conserved residues in class I cyclases and those in head-to-tail (aHT) trans-prenyl
transferases—outside the well-known DDXXD-like and NSE/DTE-like domains. In the
there is a highly conserved Thr>Gln>Asp>Tyr motif and in the cyclases,
a similar Thr>Arg>Asp>Tyr domain, these residues forming very similar,
extended H-bond networks (rmsd ~1.4 Å) that are involved in catalysis, leading
to the proposal that there are 3 key domains in both the cyclases and the aHT prenyltransferases:
The AC-domain that binds MgA and MgC; the B domain that binds MgB and leads to
pocket closure, ionization, and condensation or cyclization; and the D-domain
H-bond network, involved in H+ elimination. In aHH prenyltransferases
the overall folds and MgABC motifs are similar to those found in the cyclase
and aHT proteins, but the
full Thr>Arg/Gln>Asp>Tyr domain is absent and instead there are
Tyr/Asp or Tyr/Glu residues that bind to MgC and are highly conserved. Overall,
the results are of general interest since they show unexpected similarities
between the enzymes that produce the most diverse molecules on Earth: aHT and aHH prenyltransferases,
and terpenoid cyclases.