Literature summary for 4.3.1.7 extracted from

Nikolakakis, K.; Ohtaki, A.; Newton, K.; Chworos, A.; Sagermann, M.
Preliminary structural investigations of the Eut-L shell protein of the ethanolamine ammonia-lyase metabolosome of Escherichia coli (2009), Acta Crystallogr. Sect. F, 65, 128-132.

Cloned(Commentary)

Cloned (Comment)	Organism
cloning and overexpression of three different versions (eut-LpET151 his-tagged version (appends a cleavable 33-amino acid tag sequence to the N-terminus), Eut-L_NHIS (short His6-tagged version) and Eut-L_CHIS (short His6-tagged version)) of the protein is carried out directly from the Escherichia coli genome by selective PCR amplification. Selenomethione-derivatized proteins are obtained by growing cloned bacteria in selenomethionine-containing M9 minimal media. Protein overexpression and purification are performed.	Escherichia coli

Cloned (Comment)

Organism

cloning and overexpression of three different versions (eut-LpET151 his-tagged version (appends a cleavable 33-amino acid tag sequence to the N-terminus), Eut-L_NHIS (short His6-tagged version) and Eut-L_CHIS (short His6-tagged version)) of the protein is carried out directly from the Escherichia coli genome by selective PCR amplification. Selenomethione-derivatized proteins are obtained by growing cloned bacteria in selenomethionine-containing M9 minimal media. Protein overexpression and purification are performed.

Escherichia coli

Crystallization (Commentary)

Crystallization (Comment)	Organism
For crystallization trials, protein sample is dialyzed against 50 mM HEPES with pH 7.0 and concentrates to a final concentration of about 1 mg/ml. Purification results in highly pure protein that crystallizes readily under many different conditions, protein forms thin hexagonal plate-shaped crystals belonging to space group P3. Best crystals of Eut-L_NHIS are obtained in 3.3 M ammonium acetate, 5% polyethylene glycol 400 and 50 mM Tris buffer pH 7.5, crystals grow as hexagonal plates. Eut-L_CHIS crystals grow as single hexagonal plates with sharp edges. Crystals grow in 2 M NaCl, 100 mM phosphate, 100 mM MES buffer pH 6.5 and 4% PEG 400.	Escherichia coli

Crystallization (Comment)

Organism

For crystallization trials, protein sample is dialyzed against 50 mM HEPES with pH 7.0 and concentrates to a final concentration of about 1 mg/ml. Purification results in highly pure protein that crystallizes readily under many different conditions, protein forms thin hexagonal plate-shaped crystals belonging to space group P3. Best crystals of Eut-L_NHIS are obtained in 3.3 M ammonium acetate, 5% polyethylene glycol 400 and 50 mM Tris buffer pH 7.5, crystals grow as hexagonal plates. Eut-L_CHIS crystals grow as single hexagonal plates with sharp edges. Crystals grow in 2 M NaCl, 100 mM phosphate, 100 mM MES buffer pH 6.5 and 4% PEG 400.

Escherichia coli

General Stability

General Stability	Organism
unusually high stability against different solvent conditions with respect to changes in pH and ionic strength.	Escherichia coli

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
additional information	microcompartiment shell protein	Escherichia coli	-	-

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
66800	-	for the trimer, determination by gel filtration	Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ethanolamine	Escherichia coli	Escherichia coli has evolved specialized organelles (microcompartments) for the degradation of small molecular compounds such as ethanolamine and propanediol	acetaldehyde + NH3	-	?
additional information	Escherichia coli	ethanolamine ammonia-lyase microcompartment is composed of five different shell proteins that have been proposed to assemble into symmetrically shaped polyhedral particles of varying sizes	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
by nickel-affinity chromatography	Escherichia coli

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ethanolamine	Escherichia coli has evolved specialized organelles (microcompartments) for the degradation of small molecular compounds such as ethanolamine and propanediol	Escherichia coli	acetaldehyde + NH3	-	?
additional information	ethanolamine ammonia-lyase microcompartment is composed of five different shell proteins that have been proposed to assemble into symmetrically shaped polyhedral particles of varying sizes	Escherichia coli	?	-	?

Subunits

Subunits	Comment	Organism
trimer	determination by gel filtration, freshly prepared protein oligomerizes readily into trimers in the presence or absence of 5 mM beta-mercaptoethanol, monomer consists of 219-amino-acids	Escherichia coli

Synonyms

Synonyms	Comment	Organism
ethanolamine ammonia-lyase	-	Escherichia coli
ethanolamine ammonia-lyase BMC	-	Escherichia coli
eut-L	-	Escherichia coli

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
4.6	8.5	crystals have a remarkable high stability with respect to changes in pH	Escherichia coli