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Literature summary extracted from

  • Chruszcz, M.; Zimmerman, M.D.; Wang, S.; Koclega, K.D.; Zheng, H.; Evdokimova, E.; Kudritska, M.; Cymborowski, M.; Savchenko, A.; Edwards, A.; Minor, W.
    Function-Biased Choice of Additives for Optimization of Protein Crystallization: The Case of the Putative Thioesterase PA5185 from Pseudomonas aeruginosa PAO1 (2008), Cryst. Growth Des., 8, 4054-4061.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.1.2.20 additional information PA5185 contains Asp26 Pseudomonas aeruginosa PAO1

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.2.20 expressed in Escherichia coli B834(DE3)pLysS Pseudomonas aeruginosa PAO1
3.1.2.20 native protein expressed using a modified pET-15b plasmid transformed into B834-(DE3)pLysS Escherichia coli cells Pseudomonas aeruginosa PAO1

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.2.20 crystals of Se-Met-substituted protein or of the native protein obtained by hanging-drop vapor diffusion, to 1.9 A resolution. Usage of additives (ATP, 4-(2-hydroxyethyl)-1-piperazinepropanesulfonic acid, 2-(N-morpholino)ethanesulfonic acid at pH 5.9, 3-(N-morpholino)propanesulfonic acid, HEPES at pH 7.0, sodium diphosphate, glucose-6-phosphate, nondetergent sulfobetaines 201 or 256, sodium CoA, lithium myristoyl-CoA, lithium benzoyl-CoA and lithium acetyl-CoA), yielding new crystal forms. The PA5185 protein crystallizes in four different forms. Crystal growth of PA5185 optimized by applying additives with chemical similarity to a fragment of a putative PA5185 substrate (CoA or its derivative). Application of function-biased additives can be used as a standard optimization protocol for producing improved diffraction, or new crystal forms, which may lead to better understanding of the biological functions of proteins like PA5185 Pseudomonas aeruginosa PAO1
3.1.2.20 hanging-drop vapor diffusion method Pseudomonas aeruginosa PAO1

Organism

EC Number Organism UniProt Comment Textmining
3.1.2.20 Pseudomonas aeruginosa PAO1
-
-
-
3.1.2.20 Pseudomonas aeruginosa PAO1 Q9HU04
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.2.20 immobilized metal ion affinity chromatography (Ni2+), gel filtration Pseudomonas aeruginosa PAO1

Subunits

EC Number Subunits Comment Organism
3.1.2.20 homotetramer gel filtration Pseudomonas aeruginosa PAO1
3.1.2.20 tetramer experimental data and theoretical predictions, gel filtration Pseudomonas aeruginosa PAO1

Synonyms

EC Number Synonyms Comment Organism
3.1.2.20 PA5185
-
Pseudomonas aeruginosa PAO1
3.1.2.20 thioesterase PA5185
-
Pseudomonas aeruginosa PAO1