Structural and dynamic basis of substrate permissiveness in hydroxycinnamoyltransferase (HCT).
Structural and dynamic basis of substrate permissiveness in hydroxycinnamoyltransferase (HCT).
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DOI：
10.1371/journal.pcbi.1006511
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发表时间：
2018-10
影响因子：
4.3
通讯作者：
Wang Y
中科院分区：
文献类型：
--
作者：
Chiang YC;Levsh O;Lam CK;Weng JK;Wang Y
Substrate permissiveness has long been regarded as the raw materials for the evolution of new enzymatic functions. In land plants, hydroxycinnamoyltransferase (HCT) is an essential enzyme of the phenylpropanoid metabolism. Although essential enzymes are normally associated with high substrate specificity, HCT can utilize a variety of non-native substrates. To examine the structural and dynamic basis of substrate permissiveness in this enzyme, we report the crystal structure of HCT from Selaginella moellendorffii and molecular dynamics (MD) simulations performed on five orthologous HCTs from several major lineages of land plants. Through altogether 17-μs MD simulations, we demonstrate the prevalent swing motion of an arginine handle on a submicrosecond timescale across all five HCTs, which plays a key role in native substrate recognition by these intrinsically promiscuous enzymes. Our simulations further reveal how a non-native substrate of HCT engages a binding site different from that of the native substrate and diffuses to reach the catalytic center and its co-substrate. By numerically solving the Smoluchowski equation, we show that the presence of such an alternative binding site, even when it is distant from the catalytic center, always increases the reaction rate of a given substrate. However, this increase is only significant for enzyme-substrate reactions heavily influenced by diffusion. In these cases, binding non-native substrates ‘off-center’ provides an effective rationale to develop substrate permissiveness while maintaining the native functions of promiscuous enzymes. Examples abound of enzymes that can process substrates other than their native ones. However, the structural and dynamic basis of this promiscuity remains to be fully understood. In this work, we examine HCT, an intrinsically promiscuous acyltransferase with conserved function in all land plants. We uncover the sub-microsecond swing motion of a key arginine residue facilitating the recognition of both native and non-native substrates of HCT. We also quantify the impact of an off-center binding site on the non-native reaction rate. Although our calculations were inspired by HCT, the results apply in general, i.e., for enzymes heavily influenced by diffusion, binding non-native substrates ‘off-center’, even with rather weak affinity, can accelerate non-native reactions to appreciable levels.
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DOI：
10.1107/s0907444905036693
发表时间：
2006-01-01
影响因子：
2.2
作者：
Evans, P
通讯作者：
Evans, P
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
3
作者：
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通讯作者：
Gumbart, James C.
影响因子：
5.6
作者：
Matsumura, I;Ellington, AD
通讯作者：
Ellington, AD
DOI：
10.1073/pnas.0601769103
发表时间：
2006-08-01
影响因子：
11.1
作者：
Bauer, Wolfgang R.;Nadler, Walter
通讯作者：
Nadler, Walter