Title : Improved Stability of Human CGI-58 Induced by Phosphomimetic S237E Mutation - Llamas-Garcia_2022_ACS.Omega_7_12643 |
Author(s) : Llamas-Garcia ML , Paez-Perez ED , Benitez-Cardoza CG , Montero-Moran GM , Lara-Gonzalez S |
Ref : ACS Omega , 7 :12643 , 2022 |
Abstract :
In lipolysis, the activating function of CGI-58 is regulated by its interaction with perilipin 1 (PLIN1) localized on the lipid droplet (LD), and its release is controlled by phosphorylation. Once lipolysis is stimulated by catecholamines, protein kinase A (PKA)-mediated phosphorylation enables the dissociation of the CGI-58/PLIN1 complex, thereby recruiting adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) to initiate fatty acid release. It has been shown that mouse CGI-58 mutant S239E, which mimics the phosphorylation of this residue, is able to dissociate from the CGI-58/PLIN1 complex and activate ATGL. Here, we analyze the stabilizing effect on human CGI-58 of a triple tryptophan to alanine mutant (3WA) on the LD-binding motif, as well as a quadruple mutant in which the phosphomimetic S237E substitution was introduced to the 3WA construct (3WA/S237E). We found that tryptophan residues promote wild-type (WT) protein aggregation in solution since their substitution for alanine residues favors the presence of the monomer. Our experimental data showed increased thermal stability and solubility of 3WA/S237E protein compared to the 3WA mutant. Moreover, the 3WA/S237E protein showed proper folding and a functional binding site for oleoyl-CoA. The analysis of a bioinformatic three-dimensional (3D) model suggests an intramolecular interaction between the phosphomimetic glutamic acid and a residue of the alpha/beta hydrolase core. This could explain the increased solubility and stability observed in the 3WA/S237E mutant and evidences the possible role of serine 237 phosphorylation. |
PubMedSearch : Llamas-Garcia_2022_ACS.Omega_7_12643 |
PubMedID: 35474805 |
Gene_locus related to this paper: human-ABHD5 |
Gene_locus | human-ABHD5 |
Family | CGI-58_ABHD5_ABHD4 |
Llamas-Garcia ML, Paez-Perez ED, Benitez-Cardoza CG, Montero-Moran GM, Lara-Gonzalez S (2022)
Improved Stability of Human CGI-58 Induced by Phosphomimetic S237E Mutation
ACS Omega
7 :12643
Llamas-Garcia ML, Paez-Perez ED, Benitez-Cardoza CG, Montero-Moran GM, Lara-Gonzalez S (2022)
ACS Omega
7 :12643