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Literature summary for 2.7.9.6 extracted from

  • Stogios, P.J.; Cox, G.; Spanogiannopoulos, P.; Pillon, M.C.; Waglechner, N.; Skarina, T.; Koteva, K.; Guarne, A.; Savchenko, A.; Wright, G.D.
    Rifampin phosphotransferase is an unusual antibiotic resistance kinase (2016), Nat. Commun., 7, 11343 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.9.6

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta(DE3) pLysS cells Listeria monocytogenes

Crystallization (Commentary)

Crystallization (Comment) Organism
sitting drop vapor diffusion method, using 0.1 M SPG buffer pH 6, 25% (w/v) PEG3350 Listeria monocytogenes

Protein Variants

Protein Variants Comment Organism
E297A the mutant shows increased catalytic efficiency with ATP compared to the wild type enzyme Listeria monocytogenes
E667A the mutant shows reduced catalytic efficiency with ATP compared to the wild type enzyme Listeria monocytogenes
K22A inactive Listeria monocytogenes
Q337A the mutant shows increased catalytic efficiency with ATP compared to the wild type enzyme Listeria monocytogenes
R117A the mutant shows reduced catalytic efficiency with ATP compared to the wild type enzyme Listeria monocytogenes
R666A the mutant shows reduced catalytic efficiency with ATP compared to the wild type enzyme Listeria monocytogenes
T136V the mutant shows reduced catalytic efficiency with ATP compared to the wild type enzyme Listeria monocytogenes
V368E the mutant shows increased catalytic efficiency with ATP compared to the wild type enzyme Listeria monocytogenes
V368T the mutant shows increased catalytic efficiency with ATP compared to the wild type enzyme Listeria monocytogenes
Y351F the mutant shows increased catalytic efficiency with ATP compared to the wild type enzyme Listeria monocytogenes

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00057
-
 rifampicin wild type enzyme, at pH 7.5 and 25°C Listeria monocytogenes
0.00365
-
 rifampicin mutant enzyme V368T, at pH 7.5 and 25°C Listeria monocytogenes
0.00408
-
 rifampicin mutant enzyme, Y351F, at pH 7.5 and 25°C Listeria monocytogenes
0.00456
-
 rifampicin mutant enzyme V368E, at pH 7.5 and 25°C Listeria monocytogenes
0.00476
-
 rifampicin mutant enzyme E297A, at pH 7.5 and 25°C Listeria monocytogenes
0.006
-
 rifampicin mutant enzyme Q337A, at pH 7.5 and 25°C Listeria monocytogenes
0.00838
-
 ATP wild type enzyme, at pH 7.5 and 25°C Listeria monocytogenes
0.01803
-
 ATP mutant enzyme Q337A, at pH 7.5 and 25°C Listeria monocytogenes
0.02465
-
 ATP mutant enzyme E667A, at pH 7.5 and 25°C Listeria monocytogenes
0.03041
-
 ATP mutant enzyme V368T, at pH 7.5 and 25°C Listeria monocytogenes
0.03142
-
 ATP mutant enzyme, Y351F, at pH 7.5 and 25°C Listeria monocytogenes
0.03279
-
 ATP mutant enzyme V368E, at pH 7.5 and 25°C Listeria monocytogenes
0.04949
-
 ATP mutant enzyme E297A, at pH 7.5 and 25°C Listeria monocytogenes
0.05388
-
 ATP mutant enzyme T136V, at pH 7.5 and 25°C Listeria monocytogenes
0.05612
-
 ATP mutant enzyme R117A, at pH 7.5 and 25°C Listeria monocytogenes
0.16929
-
 ATP mutant enzyme R666A, at pH 7.5 and 25°C Listeria monocytogenes

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Listeria monocytogenes

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + rifampicin + H2O Listeria monocytogenes
-
 AMP + 21-phosphorifampicin + phosphate
-
 ?
ATP + rifampicin + H2O Listeria monocytogenes 4b / F2365
-
 AMP + 21-phosphorifampicin + phosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Listeria monocytogenes Q9RLT9
-
-
Listeria monocytogenes 4b / F2365 Q9RLT9
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography and Superdex 75 gel filtration Listeria monocytogenes

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + rifampicin + H2O
-
 Listeria monocytogenes AMP + 21-phosphorifampicin + phosphate
-
 ?
ATP + rifampicin + H2O
-
 Listeria monocytogenes 4b / F2365 AMP + 21-phosphorifampicin + phosphate
-
 ?

Synonyms

Synonyms Comment Organism
rifampin phosphotransferase
-
 Listeria monocytogenes
RPH
-
 Listeria monocytogenes

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.78
-
 ATP mutant enzyme E667A, at pH 7.5 and 25°C Listeria monocytogenes
2.1
-
 ATP mutant enzyme T136V, at pH 7.5 and 25°C Listeria monocytogenes
2.25
-
 rifampicin wild type enzyme, at pH 7.5 and 25°C Listeria monocytogenes
2.39
-
 ATP mutant enzyme R117A, at pH 7.5 and 25°C Listeria monocytogenes
2.71
-
 ATP mutant enzyme R666A, at pH 7.5 and 25°C Listeria monocytogenes
2.87
-
 ATP wild type enzyme, at pH 7.5 and 25°C Listeria monocytogenes
12.91
-
 ATP mutant enzyme, Y351F, at pH 7.5 and 25°C Listeria monocytogenes
15.35
-
 ATP mutant enzyme Q337A, at pH 7.5 and 25°C Listeria monocytogenes
15.37
-
 rifampicin mutant enzyme Q337A, at pH 7.5 and 25°C Listeria monocytogenes
15.9
-
 rifampicin mutant enzyme, Y351F, at pH 7.5 and 25°C Listeria monocytogenes
30.02
-
 rifampicin mutant enzyme V368T, at pH 7.5 and 25°C Listeria monocytogenes
33.53
-
 ATP mutant enzyme V368T, at pH 7.5 and 25°C Listeria monocytogenes
39.9
-
 ATP mutant enzyme V368E, at pH 7.5 and 25°C Listeria monocytogenes
43.21
-
 ATP mutant enzyme E297A, at pH 7.5 and 25°C Listeria monocytogenes
45
-
 rifampicin mutant enzyme E297A, at pH 7.5 and 25°C Listeria monocytogenes
45.72
-
 rifampicin mutant enzyme V368E, at pH 7.5 and 25°C Listeria monocytogenes

General Information

General Information Comment Organism
physiological function the enzyme confers antibiotic resistance against rifampin Listeria monocytogenes

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
16
-
 ATP mutant enzyme R666A, at pH 7.5 and 25°C Listeria monocytogenes
39.5
-
 ATP mutant enzyme T136V, at pH 7.5 and 25°C Listeria monocytogenes
43
-
 ATP mutant enzyme R117A, at pH 7.5 and 25°C Listeria monocytogenes
72
-
 ATP mutant enzyme E667A, at pH 7.5 and 25°C Listeria monocytogenes
342
-
 ATP wild type enzyme, at pH 7.5 and 25°C Listeria monocytogenes
410
-
 ATP mutant enzyme, Y351F, at pH 7.5 and 25°C Listeria monocytogenes
850
-
 ATP mutant enzyme Q337A, at pH 7.5 and 25°C Listeria monocytogenes
873
-
 ATP mutant enzyme E297A, at pH 7.5 and 25°C Listeria monocytogenes
1100
-
 ATP mutant enzyme V368T, at pH 7.5 and 25°C Listeria monocytogenes
1210
-
 ATP mutant enzyme V368E, at pH 7.5 and 25°C Listeria monocytogenes
2700
-
 rifampicin mutant enzyme Q337A, at pH 7.5 and 25°C Listeria monocytogenes
3890
-
 rifampicin mutant enzyme, Y351F, at pH 7.5 and 25°C Listeria monocytogenes
3940
-
 rifampicin wild type enzyme, at pH 7.5 and 25°C Listeria monocytogenes
8220
-
 rifampicin mutant enzyme V368T, at pH 7.5 and 25°C Listeria monocytogenes
9430
-
 rifampicin mutant enzyme E297A, at pH 7.5 and 25°C Listeria monocytogenes
9800
-
 rifampicin mutant enzyme V368E, at pH 7.5 and 25°C Listeria monocytogenes