Literature summary for 6.1.1.15 extracted from

Kamtekar, S.; Kennedy, W.D.; Wang, J.; Stathopoulos, C.; Soll, D.; Steitz, T.A.
The structural basis of cysteine aminoacylation of tRNAPro by prolyl-tRNA synthetases (2003), Proc. Natl. Acad. Sci. USA, 100, 1673-1678.

Cloned(Commentary)

Cloned (Comment)	Organism
expression as His-tagged enzyme in Escherichia coli	Methanothermobacter thermautotrophicus

Crystallization (Comment)	Organism
vapour diffusion method, equal volumes of 3.8 mg/ml protein solution and precipitant solution containing 20 mM Tris, pH 7.5, 5 mM 2-mercaptoethanol, 50-250 mM MgCl2, 20°C, X-ray diffraction structure determination at 2.55-3.1 A, and analysis	Methanothermobacter thermautotrophicus
vapour-diffusion method, protein solution is mixed with an equal volume of well solution containing 0.2 M ammonium sulfate, 20-30% PEG 4000, 5 mM 2-mercaptoethanol, 0.1 M sodium acetate, pH 4.5, 12-25°C, a few days, X-ray diffraction structure determination at 3.2 A resolution, and analysis	Methanocaldococcus jannaschii

Inhibitors	Comment	Organism	Structure
cysteinyl-sulfamoyl-adenylate	i.e. Cys-AMS, intermediate analogue, competitive inhibition	Methanothermobacter thermautotrophicus
prolyl-sulfamoyl-adenylate	i.e. Pro-AMS, intermediate analogue, competitive inhibition	Methanothermobacter thermautotrophicus

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.05	-	L-cysteine	ATP-diphosphate exchange, pH 7.5, 35°C	Methanothermobacter thermautotrophicus
0.067	-	L-proline	ATP-diphosphate exchange reaction, pH 7.5, 35°C	Methanothermobacter thermautotrophicus
0.09	-	L-cysteine	ATP-diphosphate exchange reaction, pH 7.5, 35°C	Methanocaldococcus jannaschii
0.285	-	L-proline	ATP-diphosphate exchange reaction, pH 7.5, 35°C	Methanocaldococcus jannaschii

Metals/Ions	Comment	Organism	Structure
Zn2+	chelated by 4 cysteine residues in the C-terminus	Methanothermobacter thermautotrophicus
Zn2+	chelated by 4 cysteine residues in the C-terminus	Methanocaldococcus jannaschii

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + L-proline + tRNAPro	Methanothermobacter thermautotrophicus	-	AMP + diphosphate + L-prolyl-tRNAPro	-	?
ATP + L-proline + tRNAPro	Methanocaldococcus jannaschii	-	AMP + diphosphate + L-prolyl-tRNAPro	-	?

Organism	UniProt	Comment	Textmining
Methanocaldococcus jannaschii	-	-	-
Methanothermobacter thermautotrophicus	-	gene proS	-

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli	Methanothermobacter thermautotrophicus
recombinant His-tagged enzyme from expression in Escherichia coli	Methanocaldococcus jannaschii

Reaction	Comment	Organism	Reaction ID
ATP + L-proline + tRNAPro = AMP + diphosphate + L-prolyl-tRNAPro	dual substrate specificity	Methanothermobacter thermautotrophicus	a

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + L-cysteine + tRNACys	-	Methanothermobacter thermautotrophicus	AMP + diphosphate + L-cysteinyl-tRNACys	-	?
ATP + L-proline + tRNAPro	-	Methanothermobacter thermautotrophicus	AMP + diphosphate + L-prolyl-tRNAPro	-	?
ATP + L-proline + tRNAPro	-	Methanocaldococcus jannaschii	AMP + diphosphate + L-prolyl-tRNAPro	-	?
additional information	the enzyme also performs the ATP-diphosphate exchange reaction	Methanothermobacter thermautotrophicus	?	-	?

Synonyms	Comment	Organism
Global RNA synthesis factor	-	Methanothermobacter thermautotrophicus
Global RNA synthesis factor	-	Methanocaldococcus jannaschii
Pro-tRNA synthetase	-	Methanothermobacter thermautotrophicus
Pro-tRNA synthetase	-	Methanocaldococcus jannaschii
Proline translase	-	Methanothermobacter thermautotrophicus
Proline translase	-	Methanocaldococcus jannaschii
Proline--tRNA ligase	-	Methanothermobacter thermautotrophicus
Proline--tRNA ligase	-	Methanocaldococcus jannaschii
Prolyl RNA synthetase	-	Methanothermobacter thermautotrophicus
Prolyl RNA synthetase	-	Methanocaldococcus jannaschii
Prolyl-transfer ribonucleate synthetase	-	Methanothermobacter thermautotrophicus
Prolyl-transfer ribonucleate synthetase	-	Methanocaldococcus jannaschii
Prolyl-transfer ribonucleic acid synthetase	-	Methanothermobacter thermautotrophicus
Prolyl-transfer ribonucleic acid synthetase	-	Methanocaldococcus jannaschii
Prolyl-transfer RNA synthetase	-	Methanothermobacter thermautotrophicus
Prolyl-transfer RNA synthetase	-	Methanocaldococcus jannaschii
Prolyl-tRNA synthetase	-	Methanothermobacter thermautotrophicus
Prolyl-tRNA synthetase	-	Methanocaldococcus jannaschii
ProRS	-	Methanothermobacter thermautotrophicus
ProRS	-	Methanocaldococcus jannaschii

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
35	-	assay at	Methanothermobacter thermautotrophicus
35	-	assay at	Methanocaldococcus jannaschii

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Methanothermobacter thermautotrophicus
7.5	-	assay at	Methanocaldococcus jannaschii

Cofactor	Comment	Organism	Structure
ATP	-	Methanothermobacter thermautotrophicus
ATP	-	Methanocaldococcus jannaschii

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.000025	-	cysteinyl-sulfamoyl-adenylate	versus L-proline, pH 7.5, 35°C	Methanothermobacter thermautotrophicus
0.00005	-	prolyl-sulfamoyl-adenylate	versus L-proline, pH 7.5, 35°C	Methanothermobacter thermautotrophicus