Establishment of glutamine synthetase of Mycobacterium smegmatis as a protein acetyltransferase utilizing polyphenolic acetates as the acetyl group donors

Garima Gupta, Anil Singh Baghel, Seema Bansal, Tapesh Kumar Tyagi, Ranju Kumari, Neeraj K. Saini, Prija Ponnan, Ajit Kumar, Mridula Bose, Daman Saluja, Shamkant Anant Patkar, Virinder Singh Parmar, Hanumantharao Guru Raj

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Acetoxy Drug: Protein Transacetylase (TAase) mediating the transfer of acetyl group(s) from polyphenolic acetates (PA) to certain functional proteins in mammalian cells was identified by our earlier investigations. TAase activity was characterized in the cell lysates of Mycobacterium smegmatis and the purified protein was found to have Mr 58,000. TAase catalysed protein acetylation by a model acetoxy drug 7,8-diacetoxy-4-methylcoumarin (DAMC) was established by the demonstration of immunoreactivity of the acetylated target protein with an anti-acetyllysine antibody. The specificity of the TAase of M. smegmatis (MTAase) to various acetoxycoumarins was found to be in the order DAMC > 7-AMC > 6-AMC > 4-AC > 3-AC > ABP. Also, the N-terminal sequence of purified MTAase was found to perfectly match with glutamine synthetase (GS) of M. smegmatis. The identity of MTAase with GS was confirmed by the observation that the purified MTAase as well as the purified recombinant GS exhibited all the properties of GS. The finding that purified Escherichia coli GS was found to have substantial TAase activity highlighted the TAase function of GS in other bacteria. These results conclusively established for the first time the protein acetyltransferase function of GS of M. smegmatis.

Original languageEnglish (US)
Pages (from-to)709-715
Number of pages7
JournalJournal of Biochemistry
Volume144
Issue number6
DOIs
StatePublished - Dec 2008
Externally publishedYes

Fingerprint

Mycobacterium smegmatis
Glutamate-Ammonia Ligase
Acetyltransferases
Acetates
Proteins
Acetylation
Pharmaceutical Preparations
Escherichia coli
Anti-Idiotypic Antibodies
Bacteria
Demonstrations
Cells
Antibodies

Keywords

  • Glutamine synthetase
  • Mycobacterium smegmatis
  • Polyphenolic acetate
  • Protein acetylation
  • Transacetylase

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Establishment of glutamine synthetase of Mycobacterium smegmatis as a protein acetyltransferase utilizing polyphenolic acetates as the acetyl group donors. / Gupta, Garima; Baghel, Anil Singh; Bansal, Seema; Tyagi, Tapesh Kumar; Kumari, Ranju; Saini, Neeraj K.; Ponnan, Prija; Kumar, Ajit; Bose, Mridula; Saluja, Daman; Patkar, Shamkant Anant; Parmar, Virinder Singh; Raj, Hanumantharao Guru.

In: Journal of Biochemistry, Vol. 144, No. 6, 12.2008, p. 709-715.

Research output: Contribution to journalArticle

Gupta, G, Baghel, AS, Bansal, S, Tyagi, TK, Kumari, R, Saini, NK, Ponnan, P, Kumar, A, Bose, M, Saluja, D, Patkar, SA, Parmar, VS & Raj, HG 2008, 'Establishment of glutamine synthetase of Mycobacterium smegmatis as a protein acetyltransferase utilizing polyphenolic acetates as the acetyl group donors', Journal of Biochemistry, vol. 144, no. 6, pp. 709-715. https://doi.org/10.1093/jb/mvn124
Gupta, Garima ; Baghel, Anil Singh ; Bansal, Seema ; Tyagi, Tapesh Kumar ; Kumari, Ranju ; Saini, Neeraj K. ; Ponnan, Prija ; Kumar, Ajit ; Bose, Mridula ; Saluja, Daman ; Patkar, Shamkant Anant ; Parmar, Virinder Singh ; Raj, Hanumantharao Guru. / Establishment of glutamine synthetase of Mycobacterium smegmatis as a protein acetyltransferase utilizing polyphenolic acetates as the acetyl group donors. In: Journal of Biochemistry. 2008 ; Vol. 144, No. 6. pp. 709-715.
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abstract = "Acetoxy Drug: Protein Transacetylase (TAase) mediating the transfer of acetyl group(s) from polyphenolic acetates (PA) to certain functional proteins in mammalian cells was identified by our earlier investigations. TAase activity was characterized in the cell lysates of Mycobacterium smegmatis and the purified protein was found to have Mr 58,000. TAase catalysed protein acetylation by a model acetoxy drug 7,8-diacetoxy-4-methylcoumarin (DAMC) was established by the demonstration of immunoreactivity of the acetylated target protein with an anti-acetyllysine antibody. The specificity of the TAase of M. smegmatis (MTAase) to various acetoxycoumarins was found to be in the order DAMC > 7-AMC > 6-AMC > 4-AC > 3-AC > ABP. Also, the N-terminal sequence of purified MTAase was found to perfectly match with glutamine synthetase (GS) of M. smegmatis. The identity of MTAase with GS was confirmed by the observation that the purified MTAase as well as the purified recombinant GS exhibited all the properties of GS. The finding that purified Escherichia coli GS was found to have substantial TAase activity highlighted the TAase function of GS in other bacteria. These results conclusively established for the first time the protein acetyltransferase function of GS of M. smegmatis.",
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AU - Gupta, Garima

AU - Baghel, Anil Singh

AU - Bansal, Seema

AU - Tyagi, Tapesh Kumar

AU - Kumari, Ranju

AU - Saini, Neeraj K.

AU - Ponnan, Prija

AU - Kumar, Ajit

AU - Bose, Mridula

AU - Saluja, Daman

AU - Patkar, Shamkant Anant

AU - Parmar, Virinder Singh

AU - Raj, Hanumantharao Guru

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N2 - Acetoxy Drug: Protein Transacetylase (TAase) mediating the transfer of acetyl group(s) from polyphenolic acetates (PA) to certain functional proteins in mammalian cells was identified by our earlier investigations. TAase activity was characterized in the cell lysates of Mycobacterium smegmatis and the purified protein was found to have Mr 58,000. TAase catalysed protein acetylation by a model acetoxy drug 7,8-diacetoxy-4-methylcoumarin (DAMC) was established by the demonstration of immunoreactivity of the acetylated target protein with an anti-acetyllysine antibody. The specificity of the TAase of M. smegmatis (MTAase) to various acetoxycoumarins was found to be in the order DAMC > 7-AMC > 6-AMC > 4-AC > 3-AC > ABP. Also, the N-terminal sequence of purified MTAase was found to perfectly match with glutamine synthetase (GS) of M. smegmatis. The identity of MTAase with GS was confirmed by the observation that the purified MTAase as well as the purified recombinant GS exhibited all the properties of GS. The finding that purified Escherichia coli GS was found to have substantial TAase activity highlighted the TAase function of GS in other bacteria. These results conclusively established for the first time the protein acetyltransferase function of GS of M. smegmatis.

AB - Acetoxy Drug: Protein Transacetylase (TAase) mediating the transfer of acetyl group(s) from polyphenolic acetates (PA) to certain functional proteins in mammalian cells was identified by our earlier investigations. TAase activity was characterized in the cell lysates of Mycobacterium smegmatis and the purified protein was found to have Mr 58,000. TAase catalysed protein acetylation by a model acetoxy drug 7,8-diacetoxy-4-methylcoumarin (DAMC) was established by the demonstration of immunoreactivity of the acetylated target protein with an anti-acetyllysine antibody. The specificity of the TAase of M. smegmatis (MTAase) to various acetoxycoumarins was found to be in the order DAMC > 7-AMC > 6-AMC > 4-AC > 3-AC > ABP. Also, the N-terminal sequence of purified MTAase was found to perfectly match with glutamine synthetase (GS) of M. smegmatis. The identity of MTAase with GS was confirmed by the observation that the purified MTAase as well as the purified recombinant GS exhibited all the properties of GS. The finding that purified Escherichia coli GS was found to have substantial TAase activity highlighted the TAase function of GS in other bacteria. These results conclusively established for the first time the protein acetyltransferase function of GS of M. smegmatis.

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