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Literature summary 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)

EC Number Cloned (Comment) Organism
2.7.6.5 expression in Escherichia coli and Mycobacterium smegmatis Mycobacterium tuberculosis
3.1.7.2 expression of fragment proteins in Escherichia coli BL21(DE3) Mycobacterium tuberculosis

Protein Variants

EC Number Protein Variants Comment Organism
2.7.6.5 C633A 20fold decrease in activity Mycobacterium tuberculosis
2.7.6.5 D632A 3.5fold decrease in activity Mycobacterium tuberculosis
2.7.6.5 D81A loss of hydrolytic activity with retention of synthesis Mycobacterium tuberculosis
2.7.6.5 G241E loss of synthetic activity and retention of hydrolysis Mycobacterium tuberculosis
2.7.6.5 H344Y loss of synthetic activity and retention of hydrolysis Mycobacterium tuberculosis
2.7.6.5 H80A loss of hydrolytic activity with retention of synthesis Mycobacterium tuberculosis
2.7.6.5 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
3.1.7.2 D81A loss of hydrolytic activity with retention of synthesis Mycobacterium tuberculosis
3.1.7.2 DElTA1-86/DELTA395-738 contains only (p)ppGpp synthesis activity, no hydrolysis activity Mycobacterium tuberculosis
3.1.7.2 DELTA182-738 fragment contains only (p)ppGpp hydrolysis activity, no synthesis activity Mycobacterium tuberculosis
3.1.7.2 DELTA395-738 frament contains both synthesis and hydrolysis activities Mycobacterium tuberculosis
3.1.7.2 G241E loss of synthetic activity and retention of hydrolysis Mycobacterium tuberculosis
3.1.7.2 H344Y loss of synthetic activity and retention of hydrolysis Mycobacterium tuberculosis
3.1.7.2 H80A loss of hydrolytic activity with retention of synthesis Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.6.5 0.33
-
 GTP full length enzyme + tRNA + ribosomes + poly(U) Mycobacterium tuberculosis
2.7.6.5 0.67
-
 GTP fragment 1-394 + tRNA Mycobacterium tuberculosis
2.7.6.5 0.67
-
 GTP fragment 1-394 + tRNA + ribosomes + poly(U) Mycobacterium tuberculosis
2.7.6.5 0.67
-
 GTP fragment 1-394 monomer Mycobacterium tuberculosis
2.7.6.5 0.7
-
 GTP full length enzyme + tRNA Mycobacterium tuberculosis
2.7.6.5 0.78
-
 GTP fragment 87-394 Mycobacterium tuberculosis
2.7.6.5 0.78
-
 GTP fragment 87-394 + tRNA Mycobacterium tuberculosis
2.7.6.5 0.78
-
 GTP fragment 87-394 + tRNA + ribosomes + poly(U) Mycobacterium tuberculosis
2.7.6.5 1.36
-
 GTP full length enzyme Mycobacterium tuberculosis
3.1.7.2 additional information
-
 additional information
-
 Mycobacterium tuberculosis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.6.5 Mg2+ the optimal concentration for pppGpp synthesis is approximately equal to the total concentration of nucleotide substrates([ATP] + [GTP]) Mycobacterium tuberculosis
2.7.6.5 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.6.5 82000
-
 full-length enzyme, calculated from OD280 Mycobacterium tuberculosis
2.7.6.5 240000
-
 full-length enzyme, gel filtration Mycobacterium tuberculosis
2.7.6.5 245000
-
 full-length enzyme, calculated from amino acid sequence Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
2.7.6.5 Mycobacterium tuberculosis
-
-
-
3.1.7.2 Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.6.5
-
 Mycobacterium tuberculosis
3.1.7.2 fragments of the relMtb protein Mycobacterium tuberculosis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.6.5 0.00021
-
 mol of PPi/s/mol of enzyme, fragment 1-181 Mycobacterium tuberculosis
2.7.6.5 0.027
-
 mol of PPi/s/mol of enzyme, fragment 1-394 monomer Mycobacterium tuberculosis
2.7.6.5 0.028
-
 mol of PPi/s/mol of enzyme, full length enzyme Mycobacterium tuberculosis
2.7.6.5 0.03
-
 mol of PPi/s/mol of enzyme, fragment 1-203 Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.6.5 ATP + GTP
-
 Mycobacterium tuberculosis AMP + guanosine 5'-triphosphate 3'-diphosphate
-
 r
3.1.7.2 pppGpp + H2O
-
 Mycobacterium tuberculosis pppG + diphosphate
-
 r

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
2.7.6.5 (p)ppGpp synthetase/hydrolase
-
 Mycobacterium tuberculosis
2.7.6.5 RelMtb protein
-
 Mycobacterium tuberculosis
3.1.7.2 (p)ppGpp synthetase/hydrolase
-
 Mycobacterium tuberculosis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.6.5 0.4
-
 GTP full length enzyme Mycobacterium tuberculosis
2.7.6.5 0.4
-
 GTP full length enzyme + tRNA Mycobacterium tuberculosis
2.7.6.5 0.47
-
 GTP fragment 87-394 Mycobacterium tuberculosis
2.7.6.5 0.47
-
 GTP fragment 87-394 + tRNA Mycobacterium tuberculosis
2.7.6.5 0.47
-
 GTP fragment 87-394 + tRNA + ribosomes + poly(U) Mycobacterium tuberculosis
2.7.6.5 0.66
-
 GTP fragment 1-394 + tRNA Mycobacterium tuberculosis
2.7.6.5 0.66
-
 GTP fragment 1-394 + tRNA + ribosomes + poly(U) Mycobacterium tuberculosis
2.7.6.5 0.66
-
 GTP fragment 1-394 monomer Mycobacterium tuberculosis
2.7.6.5 8.4
-
 GTP full length enzyme + tRNA + ribosomes + poly(U) Mycobacterium tuberculosis