Literature summary extracted from

Johnson, J.M.; Sanford, B.L.; Strom, A.M.; Tadayon, S.N.; Lehman, B.P.; Zirbes, A.M.; Bhattacharyya, S.; Musier-Forsyth, K.; Hati, S.
Multiple pathways promote dynamical coupling between catalytic domains in Escherichia coli prolyl-tRNA synthetase (2013), Biochemistry, 52, 4399-4412.

Cloned(Commentary)

EC Number	Cloned (Comment)	Organism
6.1.1.15	expressed in Escherichia coli SG13009 (pREP4) cells	Escherichia coli

Protein Variants

EC Number	Protein Variants	Comment	Organism
6.1.1.15	D198A	the overall aminoacylation activity of the mutant is reduced 5.5fold	Escherichia coli
6.1.1.15	E218A/N305A	inactive	Escherichia coli
6.1.1.15	E234A	the overall aminoacylation activity of the mutant is reduced 2fold	Escherichia coli
6.1.1.15	F415A	the aminoacylation activity of the mutant is nearly abolished with rates 70fold slower than the wild type	Escherichia coli
6.1.1.15	G412A	the overall aminoacylation activity of the mutant is reduced 7fold	Escherichia coli
6.1.1.15	H302A	the overall aminoacylation activity of the mutant is reduced 2fold	Escherichia coli
6.1.1.15	H302A/G412A	the overall aminoacylation activity of the mutant is reduced 5.5fold	Escherichia coli
6.1.1.15	N305A	the aminoacylation activity of the mutant is nearly abolished with rates 70fold slower than the wild type	Escherichia coli
6.1.1.15	N305A/G412A	inactive	Escherichia coli

KM Value [mM]

EC Number	KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
6.1.1.15	0.18	-	L-proline	wild type enzyme, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	0.22	-	L-proline	mutant enzyme H302A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	0.3	-	L-proline	mutant enzyme G412A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	0.33	-	L-proline	mutant enzyme D198A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	0.45	-	L-proline	mutant enzyme N305A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	0.62	-	L-proline	mutant enzyme H302A/G412A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	0.76	-	L-proline	mutant enzyme F415A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	1.03	-	L-proline	mutant enzyme E234A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	3.4	-	L-proline	mutant enzyme E218A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number	Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
6.1.1.15	ATP + L-proline + tRNAPro	Escherichia coli	-	AMP + diphosphate + L-prolyl-tRNAPro	-	r

Organism

EC Number	Organism	UniProt	Comment	Textmining
6.1.1.15	Escherichia coli	-	-	-

Purification (Commentary)

EC Number	Purification (Comment)	Organism
6.1.1.15	Talon cobalt affinity resin column chromatography, and gel filtration	Escherichia coli

Substrates and Products (Substrate)

EC Number	Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
6.1.1.15	ATP + L-proline + tRNAPro	-	Escherichia coli	AMP + diphosphate + L-prolyl-tRNAPro	-	r

Synonyms

EC Number	Synonyms	Comment	Organism
6.1.1.15	Prolyl-tRNA synthetase	-	Escherichia coli
6.1.1.15	ProRS	-	Escherichia coli

Turnover Number [1/s]

EC Number	Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
6.1.1.15	0.131	-	L-proline	mutant enzyme F415A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	0.61	-	L-proline	mutant enzyme N305A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	4.4	-	L-proline	mutant enzyme E218A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	6.7	-	L-proline	mutant enzyme E234A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	6.98	-	L-proline	mutant enzyme D198A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	7.3	-	L-proline	mutant enzyme H302A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	10.7	-	L-proline	mutant enzyme H302A/G412A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	12.6	-	L-proline	wild type enzyme, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	12.8	-	L-proline	mutant enzyme G412A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli

Cofactor

EC Number	Cofactor	Comment	Organism	Structure
6.1.1.15	ATP	-	Escherichia coli

kcat/KM [mM/s]

EC Number	kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
6.1.1.15	0.17	-	L-proline	mutant enzyme F415A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	1.3	-	L-proline	mutant enzyme E218A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	1.4	-	L-proline	mutant enzyme N305A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	6.5	-	L-proline	mutant enzyme E234A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	17	-	L-proline	mutant enzyme H302A/G412A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	21	-	L-proline	mutant enzyme D198A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	33	-	L-proline	mutant enzyme H302A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	43	-	L-proline	mutant enzyme G412A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.1.1.15	71	-	L-proline	wild type enzyme, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli

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Release 2023.1 (January 2023)