Severinac L

References (1)

Title : Transition metal cation inhibition of Mycobacterium tuberculosis esterase RV0045C - Bowles_2021_Protein.Sci__
Author(s) : Bowles IE , Pool EH , Lancaster BS , Lawson EK , Savas CP , Kartje ZJ , Severinac L , Cho DH , Macbeth MR , Johnson RJ , Hoops GC
Ref : Protein Science , : , 2021
Abstract : Mycobacterium tuberculosis virulence is highly metal-dependent with metal availability modulating the shift from the dormant to active states of M. tuberculosis infection. Rv0045c from M. tuberculosis is a proposed metabolic serine hydrolase whose folded stability is dependent on divalent metal concentration. Herein, we measured the divalent metal inhibition profile of the enzymatic activity of Rv0045c and found specific divalent transition metal cations (Cu(2+) <= Zn(2+) > Ni(2+) > Co(2+) ) strongly inhibited its enzymatic activity. The metal cations bind allosterically, largely affecting values for k(cat) rather than K(M) . Removal of the artificial N-terminal 6xHis-tag did not change the metal-dependent inhibition, indicating that the allosteric inhibition site is native to Rv0045c. To isolate the site of this allosteric regulation in Rv0045c, the structures of Rv0045c were determined at 1.8 A and 2.0 A resolution in the presence and absence of Zn(2+) with each structure containing a previously unresolved dynamic loop spanning the binding pocket. Through the combination of structural analysis with and without zinc and targeted mutagenesis, this metal-dependent inhibition was traced to multiple chelating residues (H202A/E204A) on a flexible loop, suggesting dynamic allosteric regulation of Rv0045c by divalent metals. Although serine hydrolases like Rv0045c are a large and diverse enzyme superfamily, this is the first structural confirmation of allosteric regulation of their enzymatic activity by divalent metals. This article is protected by copyright. All rights reserved.
ESTHER : Bowles_2021_Protein.Sci__
PubMedSearch : Bowles_2021_Protein.Sci__
PubMedID: 33914998
Gene_locus related to this paper: myctu-RV0045C