Literature summary for 6.1.1.7 extracted from

Fukunaga, R.; Yokoyama, S.
Crystallization and preliminary X-ray crystallographic study of alanyl-tRNA synthetase from the archaeon Archaeoglobus fulgidus (2007), Acta Crystallogr. Sect. F, 63, 224-228.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of the full-length AlaRS, of the N-terminal fragment lacking the C-terminal oligomerization domain, residues 1-739, and of the C-terminal oligomerization domain of AlaRS, residues 737-906, in Escherichia coli strain BL21	Archaeoglobus fulgidus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant full-length enzyme, N-terminal domain, and C-terminal domain, hanging-drop vapour-diffusion method, mixing of 0.001 ml of protein and reservoir solution, and equilibration against 0.5 ml of reservoir solution at 20°C, for crystallization of the AlaRS-FLtRNAAla complex, tRNAAla is heated at 80°C for 5 min and is gradually cooled to room temperature for refolding, in presence of 1 mM AMP-PNP, different methods for the different protein samples are used, overview, X-ray diffraction structure determination and analysis at 3.2-3.5 A resolution	Archaeoglobus fulgidus

Crystallization (Comment)

Organism

purified recombinant full-length enzyme, N-terminal domain, and C-terminal domain, hanging-drop vapour-diffusion method, mixing of 0.001 ml of protein and reservoir solution, and equilibration against 0.5 ml of reservoir solution at 20°C, for crystallization of the AlaRS-FLtRNAAla complex, tRNAAla is heated at 80°C for 5 min and is gradually cooled to room temperature for refolding, in presence of 1 mM AMP-PNP, different methods for the different protein samples are used, overview, X-ray diffraction structure determination and analysis at 3.2-3.5 A resolution

Archaeoglobus fulgidus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Archaeoglobus fulgidus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + L-alanine + tRNAAla	Archaeoglobus fulgidus	-	AMP + diphosphate + L-alanyl-tRNAAla	-	?

Organism

Organism	UniProt	Comment	Textmining
Archaeoglobus fulgidus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant full-length AlaRS and AlaRS domains from Escherichia coli strain BL21 by anion exchange chromatography followed by hydrophobic interaction and adsorption chromatography, respectively	Archaeoglobus fulgidus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + L-alanine + tRNAAla	-	Archaeoglobus fulgidus	AMP + diphosphate + L-alanyl-tRNAAla	-	?
ATP + L-alanine + tRNAAla	the aminoacylation reaction takes place in two steps catalyzed by the same active site: the synthesis of an aminoacyladenylate as an activated intermediate from the amino acid and ATP and the transfer of the aminoacyl moiety to the 3'-terminus of the cognate tRNA to yield the aminoacyl-tRNA, the synthetic active site of AlaRS misrecognizes noncognate glycine and serine as well as recognizing the cognate alanine and produces GlytRNAAla and Ser-tRNAAla, the editing domain hydrolyzes the incorrect products GlytRNAAla and Ser-tRNAAla and thus contributes to accurate aminoacylation, three tRNA isoacceptors, tRNAAla1, tRNA and tRNAAla3	Archaeoglobus fulgidus	AMP + diphosphate + L-alanyl-tRNAAla	-	?

Subunits

Subunits	Comment	Organism
More	AlaRS consists of four parts: an N-terminal aminoacylation active-site domain, a tRNA-recognition module, an editing domain and a C-terminal oligomerization domain	Archaeoglobus fulgidus

Synonyms

Synonyms	Comment	Organism
Alanyl-tRNA synthetase	-	Archaeoglobus fulgidus
AlaRS	-	Archaeoglobus fulgidus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
65	-	assay at	Archaeoglobus fulgidus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Archaeoglobus fulgidus

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Archaeoglobus fulgidus