Home LiteratureArticle Details
PMID: 8982009 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Binding of L-branched-chain amino acids causes a conformational change in BkdR.

Journal of bacteriology ·Vol. 179 ·No. 1 ·1997-01-00 ·Pages 276-9

Madhusudhan KT, Huang N, Braswell EH, Sokatch JR

Abstract

BkdR is the positive transcriptional activator of the inducible bkd operon of Pseudomonas putida. Evidence is accumulating that L-branched-chain amino acids are the inducers of the operon, and the data obtained in this study show that they induce a conformational change in BkdR. Addition of L-branched-chain amino acids increased the susceptibility of BkdR to trypsin with the cleavage between Arg-51 and Gln-52 on the C-terminal side of the DNA-binding domain. L-Valine also caused an increased fluorescence emission intensity and produced significant changes in the circular dichroism spectrum of BkdR. Analytical ultracentrifugation confirmed earlier data obtained from gel filtration that BkdR was a tetramer with a Stokes radius of 32 +/- 3 A and an axial ratio of 2:1.

MeSH Terms
Amino Acids/chemistry Bacterial Proteins/chemistry DNA-Binding Proteins/chemistry Kinetics Leucine-Responsive Regulatory Protein Molecular Weight Peptides/chemistry Protein Binding Protein Conformation Protein Structure, Secondary Pseudomonas putida/chemistry Transcription Factors Trypsin/metabolism Valine/pharmacology
Chemicals
Amino Acids Bacterial Proteins DNA-Binding Proteins Peptides Transcription Factors Leucine-Responsive Regulatory Protein Trypsin Valine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Madhusudhan K T
Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City 73190, USA.
Huang N
Braswell E H
Sokatch J R
References (26)
26 references, click to expand
  1. Dual role of a single multienzyme complex in the oxidative decarboxylation of pyruvate and branched-chain 2-oxo acids in Bacillus subtilis.
    Biochem J. 1983 Oct 1;215(1):133-40 PMID: 6414463
  2. Isolation of rabbit liver branched chain alpha-ketoacid dehydrogenase and regulation by phosphorylation.
    J Biol Chem. 1982 Dec 10;257(23):14433-9 PMID: 7142221
  3. Molecular cloning of genes encoding branched-chain keto acid dehydrogenase of Pseudomonas putida.
    J Bacteriol. 1987 Apr;169(4):1619-25 PMID: 3549697
  4. Comparison of the amino acid sequences of the transacylase components of branched chain oxoacid dehydrogenase of Pseudomonas putida, and the pyruvate and 2-oxoglutarate dehydrogenases of Escherichia coli.
    Eur J Biochem. 1988 Sep 1;176(1):165-9 PMID: 3046941
  5. Similarity of the E1 subunits of branched-chain-oxoacid dehydrogenase from Pseudomonas putida to the corresponding subunits of mammalian branched-chain-oxoacid and pyruvate dehydrogenases.
    Eur J Biochem. 1988 Sep 15;176(2):311-7 PMID: 3416875
  6. The helix-turn-helix DNA binding motif.
    J Biol Chem. 1989 Feb 5;264(4):1903-6 PMID: 2644244
  7. Sequence analysis of the lpdV gene for lipoamide dehydrogenase of branched-chain-oxoacid dehydrogenase of Pseudomonas putida.
    Eur J Biochem. 1989 Jan 15;179(1):61-9 PMID: 2917566
  8. The tdh and serA operons of Escherichia coli: mutational analysis of the regulatory elements of leucine-responsive genes.
    J Bacteriol. 1991 Oct;173(19):5944-53 PMID: 1917830
  9. Boundary analysis in sedimentation transport experiments: a procedure for obtaining sedimentation coefficient distributions using the time derivative of the concentration profile.
    Anal Biochem. 1992 Jun;203(2):295-301 PMID: 1416025
  10. Lrp, a global regulatory protein of Escherichia coli, binds co-operatively to multiple sites and activates transcription of ilvIH.
    J Mol Biol. 1993 Jan 20;229(2):306-18 PMID: 8429549
  11. Mutations affecting the ability of Escherichia coli Lrp to bind DNA, activate transcription, or respond to leucine.
    J Bacteriol. 1993 Feb;175(4):1110-7 PMID: 8432705
  12. The TyrR protein of Escherichia coli, analysis by limited proteolysis of domain structure and ligand-mediated conformational changes.
    J Biol Chem. 1993 Mar 5;268(7):5040-7 PMID: 8444880
  13. Characterization of BkdR-DNA binding in the expression of the bkd operon of Pseudomonas putida.
    J Bacteriol. 1995 Feb;177(3):636-41 PMID: 7836297
  14. The leucine-responsive regulatory protein, a global regulator of metabolism in Escherichia coli.
    Microbiol Rev. 1994 Sep;58(3):466-90 PMID: 7968922
  15. The bkdR gene of Pseudomonas putida is required for expression of the bkd operon and encodes a protein related to Lrp of Escherichia coli.
    J Bacteriol. 1993 Jul;175(13):3934-40 PMID: 8320210
  16. A consensus sequence for binding of Lrp to DNA.
    J Bacteriol. 1995 Sep;177(17):4872-80 PMID: 7665463
  17. The effect of pH and temperature on the self-association of recombinant human interleukin-2 as studied by equilibrium sedimentation.
    Pharm Res. 1995 May;12(5):637-41 PMID: 7479545
  18. Crystal structure of the lactose operon repressor and its complexes with DNA and inducer.
    Science. 1996 Mar 1;271(5253):1247-54 PMID: 8638105
  19. Stoichiometry of BkdR to substrate DNA in Pseudomonas putida.
    Biochem Biophys Res Commun. 1996 Jun 14;223(2):315-9 PMID: 8670279
  20. Regulation of valine catabolism in Pseudomonas putida.
    J Bacteriol. 1972 Jun;110(3):1073-81 PMID: 5030618
  21. Cyclic adenosine 3',5'-monophosphate levels and activities of adenylate cyclase and cyclic adenosine 3',5'-monophosphate phosphodiesterase in Pseudomonas and Bacteroides.
    J Bacteriol. 1977 Jan;129(1):87-96 PMID: 187575
  22. Purification and characterization of branched chain alpha-keto acid dehydrogenase complex of bovine kidney.
    Proc Natl Acad Sci U S A. 1978 Oct;75(10):4881-5 PMID: 283398
  23. Cyclic adenosine 3',5'-monophosphate levels in Pseudomonas putida and Pseudomonas aeruginosa during induction and carbon catabolite repression of histidase synthesis.
    J Bacteriol. 1981 Mar;145(3):1286-92 PMID: 6259129
  24. Purification of a branched-chain keto acid dehydrogenase from Pseudomonas putida.
    J Bacteriol. 1981 Nov;148(2):647-52 PMID: 7298579
  25. Analysis of data from the analytical ultracentrifuge by nonlinear least-squares techniques.
    Biophys J. 1981 Dec;36(3):575-88 PMID: 7326325
  26. Resolution of branched-chain oxo acid dehydrogenase complex of Pseudomonas aeruginosa PAO.
    Biochem J. 1986 Feb 1;233(3):737-42 PMID: 3085653
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.1.276-279.1997
Published
1997-01-00
Pages
276-9
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC178690
Subset
IM
Grants
NIDDK NIH HHS · DK 21737 · United States
External Links
PubMed source DOI: 10.1128/jb.179.1.276-279.1997 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2