%0 Journal Article
%A Bleffert, Florian
%A Granzin, Joachim
%A Gohlke, Holger
%A Batra-Safferling, Renu
%A Jaeger, Karl-Erich
%A Kovacic, Filip
%T Pseudomonas aeruginosa esterase PA2949, a bacterial homolog of the human membrane esterase ABHD6: expression, purification and crystallization
%J Acta crystallographica / F Structural biology communications Section F
%V F75
%N 4
%@ 2053-230X
%C Oxford [u.a.]
%I Blackwell
%M FZJ-2019-02510
%P 270 - 277
%D 2019
%X 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.
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:30950828
%U <Go to ISI:>//WOS:000463600400008
%R 10.1107/S2053230X19002152
%U https://juser.fz-juelich.de/record/862156