glnA [2017-8-11 16:0:3]

trigger enzyme

locus: BSU17460

pI: 4.87

mw: 50.11 kDa

protein length: 444 aa

gene length: 1332 bp

function: glutamine biosynthesis, control of TnrA and GlnR activity

product: trigger enzyme

essential: no

ec: 6.3.1.2

synonyms

Genomic Context

categories

• [category|SW 2|Metabolism] → [category|SW 2.3|Amino acid/ nitrogen metabolism] → [category|SW 2.3.1|Biosynthesis/ acquisition of amino acids] → [category|SW 2.3.1.1|Biosynthesis/ acquisition of glutamate/ glutamine/ ammonium assimilation]
• [category|SW 3|Information processing] → [category|SW 3.4|Regulation of gene expression] → [category|SW 3.4.2|Transcription factors and their control] → [category|SW 3.4.2.7|Control of transcription factor (other than two-component system)]
• [category|SW 3|Information processing] → [category|SW 3.4|Regulation of gene expression] → [category|SW 3.4.3|Trigger enzyme] → [category|SW 3.4.3.2|Trigger enzymes that control gene expression by protein-protein interaction with transcription factors]
• [category|SW 6|Groups of genes] → [category|SW 6.4|Phosphoproteins] → [category|SW 6.4.8|Phosphorylation on either a Ser, Thr or Tyr residue]

Gene

Coordinates: 1,878,425 → 1,879,759

Phenotypes of a mutant

• auxotrophic for glutamine

The protein

Catalyzed reaction/ biological activity

• ATP L-glutamate NH₃ = ADP phosphate L-glutamine (according to Swiss-Prot)

Protein family

• glutamine synthetase family (according to Swiss-Prot)

Kinetic information

• K(M) for: Glu: 27 mM, ATP: 2.4 mM, ammonium: 0.18 mM; v(max): 3.7 µmol/min/mg

[SW|Domains]

• glutamate binding flap (aa 300 ... 306: protects unstable intermediates from abberant hydrolysis)

Modification

• phosphorylated on ser/ thr/ tyr [Pubmed|16493705]
• ''in vitro'' phosphorylated by [protein|23FF4A00C36EC1B7297F51A9EF3579B41F0B7EBA|PrkC] on Thr-26, Thr-147, Ser-207, and Thr-286 [Pubmed|20389117]

[SW|Cofactors]

• Mg(2 )

Effectors of protein activity

• feedback inhibition by glutamine, glutamine binds the entrance site for glutamate
• activity is inhibited upon interaction with [protein|857EF1AC54DEA0690C01B7F8CEFFD16312C58F20|TnrA] [Pubmed|23535029]

Structure

• [PDB|4LNN] (apo-GS) [Pubmed|24158439]
• [PDB|3QAJ] (complex with ATP)
• [http://pdb.org/pdb/search/structidSearch.do?structureId=4s0r 4S0R] (the [protein|857EF1AC54DEA0690C01B7F8CEFFD16312C58F20|TnrA]-[protein|68F4E792B99F86AE9E8F06D2200E128937331F5D|GlnA] complex) [Pubmed|25691471]
• [PDB|A general discussion of GS structure]

[SW|Localization]

• cytoplasm (according to Swiss-Prot)

Additional information

• GlnA is a homooligomer of 12 subunits
• belongs to the 100 [SW|most abundant proteins] [PubMed|15378759]

Expression and Regulation

Operons

• genes: [gene|641C4BDD9702804642E1753A9C779E80FABB3919|glnR]-[gene|68F4E792B99F86AE9E8F06D2200E128937331F5D|glnA]
  description: [Pubmed|8636055]

  sigma factors

  • [protein|360F48D576DE950DF79C1A2677B7A35A8D8CC30C|SigA]: sigma factor, [Pubmed|2906311], in [regulon|360F48D576DE950DF79C1A2677B7A35A8D8CC30C|SigA regulon]

  regulatory mechanism

  • [protein|857EF1AC54DEA0690C01B7F8CEFFD16312C58F20|TnrA]: repression, [Pubmed|8799114], in [regulon|857EF1AC54DEA0690C01B7F8CEFFD16312C58F20|TnrA regulon]
  • [protein|641C4BDD9702804642E1753A9C779E80FABB3919|GlnR]: repression, in complex with [protein|68F4E792B99F86AE9E8F06D2200E128937331F5D|GlnA], in [regulon|641C4BDD9702804642E1753A9C779E80FABB3919|GlnR regulon]

  regulation

  • expressed in the absence of glutamine ([protein|641C4BDD9702804642E1753A9C779E80FABB3919|GlnR]) [Pubmed|8636055]
  • the mRNA is processed between [gene|641C4BDD9702804642E1753A9C779E80FABB3919|glnR] and [gene|68F4E792B99F86AE9E8F06D2200E128937331F5D|glnA] by [protein|872CCB5A49C9000BD95E4B0472556D5F60F7D7A4|RNase Y], this requires the [protein|EE52DFA35B935E551871D079A9BE877DB2001A3B|YmcA]-[protein|6C9A092F38739A3759793EF8B496569CD02C2E3F|YlbF]-[protein|EBD15C174A03B7FCDFFE4C5DB5D86E93F1B9CAC4|YaaT] complex [Pubmed|29794222]

• genes: [gene|9F6B78C1932FB56F7F71430DB16BC862A312407B|ynbA]-[gene|14EFA3BF4E5DD5368EF02B0733300CEF7A4E0DC7|ynbB]-[gene|641C4BDD9702804642E1753A9C779E80FABB3919|glnR]-[gene|68F4E792B99F86AE9E8F06D2200E128937331F5D|glnA]
  description: [Pubmed|22383849]

  regulation

  • the mRNA is processed between [gene|641C4BDD9702804642E1753A9C779E80FABB3919|glnR] and [gene|68F4E792B99F86AE9E8F06D2200E128937331F5D|glnA] by [protein|872CCB5A49C9000BD95E4B0472556D5F60F7D7A4|RNase Y], this requires the [protein|EE52DFA35B935E551871D079A9BE877DB2001A3B|YmcA]-[protein|6C9A092F38739A3759793EF8B496569CD02C2E3F|YlbF]-[protein|EBD15C174A03B7FCDFFE4C5DB5D86E93F1B9CAC4|YaaT] complex [Pubmed|29794222]

• genes: [gene|68F4E792B99F86AE9E8F06D2200E128937331F5D|glnA]
  description: [Pubmed|22383849]

Biological materials

Mutant

• GP247 (''glnA::cat''), available in [SW|Jörg Stülke]'s lab
• BKE17460 (''glnA''::''ermC'') (available at the [http://www.bgsc.org/ BGSC] and in [SW|Jörg Stülke]'s lab) [pubmed|28189581]
• GP2263 (''glnA''::''ermC'') (available in [SW|Jörg Stülke]'s lab)
• BP148 (del(''[gene|641C4BDD9702804642E1753A9C779E80FABB3919|glnR]-[gene|68F4E792B99F86AE9E8F06D2200E128937331F5D|glnA]'')::''cat''), available in [SW|Fabian Commichau]'s lab
• GP1883 (del(''[gene|641C4BDD9702804642E1753A9C779E80FABB3919|glnR]-[gene|68F4E792B99F86AE9E8F06D2200E128937331F5D|glnA]'')::''ermC''), available in [SW|Fabian Commichau]'s and [SW|Jörg Stülke]'s labs

Expression vector

• expression/ purification from ''E. coli'', with N-terminal Strep-tag (in [SW|pGP172]): pGP174, available in [SW|Jörg Stülke]'s lab
• pGP177 (N-terminal Strep-tag, purification from ''B. subtilis'', for [SW|SPINE], in [SW|pBQ200]), available in [SW|Jörg Stülke]'s lab

lacZ fusion

• ''[gene|641C4BDD9702804642E1753A9C779E80FABB3919|glnR]-lacZ'': pGP189 (in [protein|search|pAC7]), available in [SW|Jörg Stülke]'s lab

Antibody

• available in [SW|Karl Forchhammer]'s lab

Labs working on this gene/protein

• [SW|Susan Fisher], Boston, USA [http://www.bumc.bu.edu/microbiology/research/susan-h-fisher-phd/ homepage]

References

Reviews

• 10231480,18086213,22625175

Original publications

• 19233925,20389117,8799114,18195355,11719184,12139611,2573733,8636055,19233925,16493705,16885465,6141156,2906311,20656908,16055443,25755103,18331450,16547045,8093698,21435182,23535029,24158439,15378759,25691471,26635369,26883633