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Literature summary for 2.7.6.5 extracted from

  • Avarbock, A.; Avarbock, D.; Teh, J.S.; Buckstein, M.; Wang, Z.m.; Rubin, H.
    Functional Regulation of the Opposing (p)ppGpp Synthetase/Hydrolase Activities of RelMtb from Mycobacterium tuberculosis (2005), Biochemistry, 44, 9913-9923.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli and Mycobacterium smegmatis Mycobacterium tuberculosis

Protein Variants

Protein Variants Comment Organism
C633A 20fold decrease in activity Mycobacterium tuberculosis
D632A 3.5fold decrease in activity Mycobacterium tuberculosis
D81A loss of hydrolytic activity with retention of synthesis Mycobacterium tuberculosis
G241E loss of synthetic activity and retention of hydrolysis Mycobacterium tuberculosis
H344Y loss of synthetic activity and retention of hydrolysis Mycobacterium tuberculosis
H80A loss of hydrolytic activity with retention of synthesis Mycobacterium tuberculosis
additional information fragments 1-203 (22.7 kDa) and 1-181 (20.1 kDa) possess hydrolytic activity but are incapable of synthesis activity, fragment 1-156 (17.3 kDa) is not capable of either synthesis or hydolytic activity, relative acticity of fragment 1-181 decreases approximately 130fold compared to that of the wild type, fragment 87-394 (35.1 kDa) has only synthesis activity, fragment 1-394 (44.6 kDa) and 1-450 are capable of synthesis and hydrolysis, the trimer state of fragment 1-394 appears to be a catalytically less efficient state than the monomer state, fragment 395-738 is devoid of any activity Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.33
-
GTP full length enzyme + tRNA + ribosomes + poly(U) Mycobacterium tuberculosis
0.67
-
GTP fragment 1-394 + tRNA Mycobacterium tuberculosis
0.67
-
GTP fragment 1-394 + tRNA + ribosomes + poly(U) Mycobacterium tuberculosis
0.67
-
GTP fragment 1-394 monomer Mycobacterium tuberculosis
0.7
-
GTP full length enzyme + tRNA Mycobacterium tuberculosis
0.78
-
GTP fragment 87-394 Mycobacterium tuberculosis
0.78
-
GTP fragment 87-394 + tRNA Mycobacterium tuberculosis
0.78
-
GTP fragment 87-394 + tRNA + ribosomes + poly(U) Mycobacterium tuberculosis
1.36
-
GTP full length enzyme Mycobacterium tuberculosis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ the optimal concentration for pppGpp synthesis is approximately equal to the total concentration of nucleotide substrates([ATP] + [GTP]) Mycobacterium tuberculosis
Mn2+ the optimal concentration for pppGpp synthesis is approximately one-half of the total concentration of nucleotide substrates([ATP] + [GTP]) Mycobacterium tuberculosis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
82000
-
full-length enzyme, calculated from OD280 Mycobacterium tuberculosis
240000
-
full-length enzyme, gel filtration Mycobacterium tuberculosis
245000
-
full-length enzyme, calculated from amino acid sequence Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + GTP Mycobacterium tuberculosis
-
AMP + guanosine 5'-triphosphate 3'-diphosphate
-
r

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Mycobacterium tuberculosis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.00021
-
mol of PPi/s/mol of enzyme, fragment 1-181 Mycobacterium tuberculosis
0.027
-
mol of PPi/s/mol of enzyme, fragment 1-394 monomer Mycobacterium tuberculosis
0.028
-
mol of PPi/s/mol of enzyme, full length enzyme Mycobacterium tuberculosis
0.03
-
mol of PPi/s/mol of enzyme, fragment 1-203 Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + GTP
-
Mycobacterium tuberculosis AMP + guanosine 5'-triphosphate 3'-diphosphate
-
r

Subunits

Subunits Comment Organism
trimer 3 * 82000, gel filtration, calculated from OD280 Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
(p)ppGpp synthetase/hydrolase
-
Mycobacterium tuberculosis
RelMtb protein
-
Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.4
-
GTP full length enzyme Mycobacterium tuberculosis
0.4
-
GTP full length enzyme + tRNA Mycobacterium tuberculosis
0.47
-
GTP fragment 87-394 Mycobacterium tuberculosis
0.47
-
GTP fragment 87-394 + tRNA Mycobacterium tuberculosis
0.47
-
GTP fragment 87-394 + tRNA + ribosomes + poly(U) Mycobacterium tuberculosis
0.66
-
GTP fragment 1-394 + tRNA Mycobacterium tuberculosis
0.66
-
GTP fragment 1-394 + tRNA + ribosomes + poly(U) Mycobacterium tuberculosis
0.66
-
GTP fragment 1-394 monomer Mycobacterium tuberculosis
8.4
-
GTP full length enzyme + tRNA + ribosomes + poly(U) Mycobacterium tuberculosis