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000862156 1001_ $$0P:(DE-Juel1)159575$$aBleffert, Florian$$b0$$ufzj
000862156 245__ $$aPseudomonas aeruginosa esterase PA2949, a bacterial homolog of the human membrane esterase ABHD6: expression, purification and crystallization
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000862156 520__ $$aThe 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.
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000862156 7001_ $$0P:(DE-Juel1)131965$$aGranzin, Joachim$$b1$$ufzj
000862156 7001_ $$0P:(DE-Juel1)172663$$aGohlke, Holger$$b2$$ufzj
000862156 7001_ $$0P:(DE-Juel1)131950$$aBatra-Safferling, Renu$$b3$$eCorresponding author$$ufzj
000862156 7001_ $$0P:(DE-Juel1)131457$$aJaeger, Karl-Erich$$b4$$eCorresponding author$$ufzj
000862156 7001_ $$0P:(DE-Juel1)131480$$aKovacic, Filip$$b5$$eCorresponding author
000862156 773__ $$0PERI:(DE-600)2175956-X$$a10.1107/S2053230X19002152$$gVol. 75, no. 4, p. 270 - 277$$n4$$p270 - 277$$tActa crystallographica / F Structural biology communications Section F$$vF75$$x2053-230X$$y2019
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