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PMID: 17190799 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Inhibitory complex of the transmembrane ammonia channel, AmtB, and the cytosolic regulatory protein, GlnK, at 1.96 A.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 104 ·No. 1 ·2007-01-02 ·Pages 42-7

Gruswitz F, O'Connell J, Stroud RM

Abstract

Ammonia conductance is highly regulated. A P(II) signal transduction protein, GlnK, is the final regulator of transmembrane ammonia conductance by the ammonia channel AmtB in Escherichia coli. The complex formed between AmtB and inhibitory GlnK at 1.96-A resolution shows that the trimeric channel is blocked directly by GlnK and how, in response to intracellular nitrogen status, the ability of GlnK to block the channel is regulated by uridylylation/deuridylylation at Y51. ATP and Mg(2+) augment the interaction of GlnK. The hydrolyzed product, adenosine 5'-diphosphate orients the surface of GlnK for AmtB blockade. 2-Oxoglutarate diminishes AmtB/GlnK association, and sites for 2-oxoglutarate are evaluated.

MeSH Terms
Adenosine Triphosphate/metabolism Cation Transport Proteins/antagonists & inhibitors,chemistry Escherichia coli Proteins/antagonists & inhibitors,chemistry,physiology Ketoglutaric Acids/metabolism Nucleotidyltransferases/chemistry,physiology PII Nitrogen Regulatory Proteins/chemistry,physiology Periplasm/metabolism Protein Conformation Protein Processing, Post-Translational Quaternary Ammonium Compounds/metabolism
Chemicals
AmtB protein, E coli Cation Transport Proteins Escherichia coli Proteins Ketoglutaric Acids PII Nitrogen Regulatory Proteins Quaternary Ammonium Compounds Adenosine Triphosphate glnK protein, E coli Nucleotidyltransferases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Gruswitz Franz
Department of Biochemistry and Biophysics, Genentech Hall, School of Medicine, University of California, 600 16th Street, San Francisco, CA 94158-2517, USA.
O'Connell Joseph
Stroud Robert M
References (25)
25 references, click to expand
  1. Phosphorylation of the signal transducer PII protein and an additional effector are required for the PII-mediated regulation of nitrate and nitrite uptake in the Cyanobacterium synechococcus sp. PCC 7942.
    Eur J Biochem. 2000 Jan;267(2):591-600 PMID: 10632730
  2. The glnKamtB operon. A conserved gene pair in prokaryotes.
    Trends Genet. 2000 Jan;16(1):11-4 PMID: 10637624
  3. Structure-function relationships of glutamine synthetases.
    Biochim Biophys Acta. 2000 Mar 7;1477(1-2):122-45 PMID: 10708854
  4. Membrane topology of the Mep/Amt family of ammonium transporters.
    Mol Microbiol. 2000 Jul;37(2):331-44 PMID: 10931328
  5. The structure of the PII-ATP complex.
    Eur J Biochem. 2001 Apr;268(7):2028-37 PMID: 11277925
  6. DOCK 4.0: search strategies for automated molecular docking of flexible molecule databases.
    J Comput Aided Mol Des. 2001 May;15(5):411-28 PMID: 11394736
  7. Molecular mechanism of acute ammonia toxicity: role of NMDA receptors.
    Neurochem Int. 2002 Aug-Sep;41(2-3):95-102 PMID: 12020609
  8. Herbaspirillum seropedicae signal transduction protein PII is structurally similar to the enteric GlnK.
    Eur J Biochem. 2002 Jul;269(13):3296-303 PMID: 12084071
  9. The structures of the PII proteins from the cyanobacteria Synechococcus sp. PCC 7942 and Synechocystis sp. PCC 6803.
    Acta Crystallogr D Biol Crystallogr. 2003 Dec;59(Pt 12):2183-90 PMID: 14646076
  10. Ammonium sensing in Escherichia coli. Role of the ammonium transporter AmtB and AmtB-GlnK complex formation.
    J Biol Chem. 2004 Mar 5;279(10):8530-8 PMID: 14668330
  11. Mechanism of ammonia transport by Amt/MEP/Rh: structure of AmtB at 1.35 A.
    Science. 2004 Sep 10;305(5690):1587-94 PMID: 15361618
  12. Mutant strains (nit) of Salmonella typhimurium with a pleiotropic defect in nitrogen metabolism.
    J Bacteriol. 1976 Oct;128(1):86-98 PMID: 10275
  13. Ammonium transport in the kidney.
    Physiol Rev. 1989 Jan;69(1):179-249 PMID: 2643123
  14. Structure of the Escherichia coli signal transducing protein PII.
    Structure. 1994 Oct 15;2(10):981-90 PMID: 7866749
  15. The Escherichia coli PII signal transduction protein is activated upon binding 2-ketoglutarate and ATP.
    J Biol Chem. 1995 Jul 28;270(30):17797-807 PMID: 7629080
  16. GlnK, a PII-homologue: structure reveals ATP binding site and indicates how the T-loops may be involved in molecular recognition.
    J Mol Biol. 1998 Sep 11;282(1):149-65 PMID: 9733647
  17. Phylogenetic characterization of novel transport protein families revealed by genome analyses.
    Biochim Biophys Acta. 1999 Feb 25;1422(1):1-56 PMID: 10082980
  18. The mechanism of ammonia transport based on the crystal structure of AmtB of Escherichia coli.
    Proc Natl Acad Sci U S A. 2004 Dec 7;101(49):17090-5 PMID: 15563598
  19. Complex formation and catalytic activation by the PII signaling protein of N-acetyl-L-glutamate kinase from Synechococcus elongatus strain PCC 7942.
    J Biol Chem. 2004 Dec 31;279(53):55202-10 PMID: 15502156
  20. Crystal structures of the signal transducing protein GlnK from Thermus thermophilus HB8.
    J Struct Biol. 2005 Jan;149(1):99-110 PMID: 15629661
  21. Unique mechanistic features of post-translational regulation of glutamine synthetase activity in Methanosarcina mazei strain Gö1 in response to nitrogen availability.
    Mol Microbiol. 2005 Mar;55(6):1841-54 PMID: 15752204
  22. PIBASE: a comprehensive database of structurally defined protein interfaces.
    Bioinformatics. 2005 May 1;21(9):1901-7 PMID: 15657096
  23. Crystal structure of the archaeal ammonium transporter Amt-1 from Archaeoglobus fulgidus.
    Proc Natl Acad Sci U S A. 2005 Oct 18;102(42):14994-9 PMID: 16214888
  24. The Escherichia coli AmtB protein as a model system for understanding ammonium transport by Amt and Rh proteins.
    Transfus Clin Biol. 2006 Mar-Apr;13(1-2):97-102 PMID: 16563828
  25. In vitro analysis of the Escherichia coli AmtB-GlnK complex reveals a stoichiometric interaction and sensitivity to ATP and 2-oxoglutarate.
    J Biol Chem. 2006 Oct 6;281(40):29558-67 PMID: 16864585
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0609796104
Published
2007-01-02
Epub
2006-00-26
Pages
42-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC1765473
Subset
IM
Grants
NIGMS NIH HHS · R37 GM024485 · United States
NIGMS NIH HHS · R01 GM024485 · United States
NIGMS NIH HHS · GM24485 · United States
NIGMS NIH HHS · GM074929 · United States
NIGMS NIH HHS · U54 GM074929 · United States
Databases
PDB
2NS1
External Links
PubMed source DOI: 10.1073/pnas.0609796104 PMC Full Text View journal details
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