Literature summary for 6.1.1.15 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.

Search for more literature for 6.1.1.15

Cloned(Commentary)

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

Protein Variants

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

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
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
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
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
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
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
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
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
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
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)

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

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	-	-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms	Comment	Organism
Prolyl-tRNA synthetase	-	Escherichia coli
ProRS	-	Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
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
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
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
6.7	-	L-proline	mutant enzyme E234A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
6.98	-	L-proline	mutant enzyme D198A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
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
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
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
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

Cofactor	Comment	Organism	Structure
ATP	-	Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
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
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
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
6.5	-	L-proline	mutant enzyme E234A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
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
21	-	L-proline	mutant enzyme D198A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
33	-	L-proline	mutant enzyme H302A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
43	-	L-proline	mutant enzyme G412A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C	Escherichia coli
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)