This title appears in the Scientific Report : 2019 

Pseudomonas aeruginosa esterase PA2949, a bacterial homolog of the human membrane esterase ABHD6: expression, purification and crystallization
Bleffert, Florian
Granzin, Joachim / Gohlke, Holger / Batra-Safferling, Renu (Corresponding author) / Jaeger, Karl-Erich (Corresponding author) / Kovacic, Filip (Corresponding author)
Institut für Molekulare Enzymtechnologie (HHUD); IMET
Strukturbiochemie ; ICS-6
Jülich Supercomputing Center; JSC
John von Neumann - Institut für Computing; NIC
Acta crystallographica / F Structural biology communications Section F, F75 (2019) 4, S. 270 - 277
Oxford [u.a.] Blackwell 2019
10.1107/S2053230X19002152
Journal Article
Forschergruppe Gohlke
Functional Macromolecules and Complexes
Biotechnology
Computational Science and Mathematical Methods
OpenAccess
OpenAccess
OpenAccess
OpenAccess
Please use the identifier: http://dx.doi.org/10.1107/S2053230X19002152 in citations.
Please use the identifier: http://hdl.handle.net/2128/22134 in citations.
The human membrane-bound α/β-hydrolase domain 6 (ABHD6) protein modulates endocannabinoid signaling, which controls appetite, pain and learning, as well as being linked to Alzheimer's and Parkinson's diseases, through the degradation of the key lipid messenger 2-arachidonylglycerol (2-AG). This makes ABHD6 an attractive therapeutic target that lacks structural information. In order to better understand the molecular mechanism of 2-AG-hydrolyzing enzymes, the PA2949 protein from Pseudomonas aeruginosa, which has 49% sequence similarity to the ABHD6 protein, was cloned, overexpressed, purified and crystallized. Overexpression of PA2949 in the homologous host yielded the membrane-bound enzyme, which was purified in milligram amounts. Besides their sequence similarity, the enzymes both show specificity for the hydrolysis of 2-AG and esters of medium-length fatty acids. PA2949 in the presence of n-octyl β-D-glucoside showed a higher activity and stability at room temperature than those previously reported for PA2949 overexpressed and purified from Escherichia coli. A suitable expression host and stabilizing detergent were crucial for obtaining crystals, which belonged to the tetragonal space group I4122 and diffracted to a resolution of 2.54 Å. This study provides hints on the functional similarity of ABHD6-like proteins in prokaryotes and eukaryotes, and might guide the structural study of these difficult-to-crystallize proteins.