001     30967
005     20200423203530.0
024 7 _ |a pmid:12741850
|2 pmid
024 7 _ |a 10.1021/bi0340595
|2 DOI
024 7 _ |a WOS:000182934300041
|2 WOS
024 7 _ |a 2128/2417
|2 Handle
024 7 _ |a altmetric:21810231
|2 altmetric
037 _ _ |a PreJuSER-30967
041 _ _ |a eng
082 _ _ |a 570
084 _ _ |2 WoS
|a Biochemistry & Molecular Biology
100 1 _ |a Gerharz, T.
|b 0
|u FZJ
|0 P:(DE-Juel1)VDB2886
245 _ _ |a Identification of basic amino acid residues important for citrate binding by the periplasmic receptor domain of the sensor kinase CitA
260 _ _ |a Columbus, Ohio
|b American Chemical Society
|c 2003
300 _ _ |a 5917 - 5924
336 7 _ |a Journal Article
|0 PUB:(DE-HGF)16
|2 PUB:(DE-HGF)
336 7 _ |a Output Types/Journal article
|2 DataCite
336 7 _ |a Journal Article
|0 0
|2 EndNote
336 7 _ |a ARTICLE
|2 BibTeX
336 7 _ |a JOURNAL_ARTICLE
|2 ORCID
336 7 _ |a article
|2 DRIVER
440 _ 0 |a Biochemistry
|x 0006-2960
|0 798
|v 42
500 _ _ |a Record converted from VDB: 12.11.2012
520 _ _ |a The sensor kinase CitA and the response regulator CitB of Klebsiella pneumoniae form the paradigm of a subfamily of bacterial two-component regulatory systems that are capable of sensing tri- or dicarboxylates in the environment and then induce transporters for the uptake of these compounds. We recently showed that the separated periplasmic domain of CitA, termed CitAP (encompasses residues 45-176 supplemented with an N-terminal methionine residue and a C-terminal hexahistidine tag), is a highly specific citrate receptor with a K(d) of 5.5 microM at pH 7. To identify positively charged residues involved in binding the citrate anion, each of the arginine, lysine, and histidine residues in CitAP was exchanged for alanine, and the resulting 17 muteins were analyzed by isothermal titration calorimetry (ITC). In 12 cases, the K(d) for citrate was identical to that of wild-type CitAP or slightly changed (3.9-17.2 microM). In one case (R98A), the K(d) was 6-fold decreased (0.8 microM), whereas in four cases (R66A, H69A, R107A, and K109A) the K(d) was 38- to >300-fold increased (0.2 to >1 mM). The secondary structure of the latter five proteins in their apo-form as deduced from far-UV circular dichroism (CD) spectra did not differ from the apo-form of wild-type CitAP; however, all of them showed an increased thermostability. Citrate increased the melting point (T(m)) of wild-type CitAP and mutein R98A by 6.2 and 9.5 degrees C, respectively, but had no effect on the T(m) of the four proteins with disturbed binding. Three of the residues important for citrate binding (R66, H69, and R107) are highly conserved in the CitA subfamily of sensor kinases, indicating that they might be involved in ligand binding by many of these sensor kinases.
536 _ _ |a Biotechnologie
|c L02
|2 G:(DE-HGF)
|0 G:(DE-Juel1)FUEK256
|x 0
588 _ _ |a Dataset connected to Web of Science, Pubmed
650 _ 2 |2 MeSH
|a Amino Acid Sequence
650 _ 2 |2 MeSH
|a Amino Acid Substitution
650 _ 2 |2 MeSH
|a Amino Acids, Basic: chemistry
650 _ 2 |2 MeSH
|a Bacterial Proteins: chemistry
650 _ 2 |2 MeSH
|a Bacterial Proteins: genetics
650 _ 2 |2 MeSH
|a Bacterial Proteins: metabolism
650 _ 2 |2 MeSH
|a Base Sequence
650 _ 2 |2 MeSH
|a Binding Sites
650 _ 2 |2 MeSH
|a Circular Dichroism
650 _ 2 |2 MeSH
|a Citric Acid: metabolism
650 _ 2 |2 MeSH
|a DNA, Bacterial: genetics
650 _ 2 |2 MeSH
|a Escherichia coli Proteins: chemistry
650 _ 2 |2 MeSH
|a Escherichia coli Proteins: genetics
650 _ 2 |2 MeSH
|a Escherichia coli Proteins: metabolism
650 _ 2 |2 MeSH
|a Kinetics
650 _ 2 |2 MeSH
|a Klebsiella pneumoniae: genetics
650 _ 2 |2 MeSH
|a Klebsiella pneumoniae: metabolism
650 _ 2 |2 MeSH
|a Ligands
650 _ 2 |2 MeSH
|a Molecular Sequence Data
650 _ 2 |2 MeSH
|a Mutagenesis, Site-Directed
650 _ 2 |2 MeSH
|a Periplasm: metabolism
650 _ 2 |2 MeSH
|a Protein Binding
650 _ 2 |2 MeSH
|a Protein Kinases: chemistry
650 _ 2 |2 MeSH
|a Protein Kinases: genetics
650 _ 2 |2 MeSH
|a Protein Kinases: metabolism
650 _ 2 |2 MeSH
|a Protein Structure, Tertiary
650 _ 2 |2 MeSH
|a Sequence Homology, Amino Acid
650 _ 2 |2 MeSH
|a Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
650 _ 2 |2 MeSH
|a Thermodynamics
650 _ 2 |2 MeSH
|a Transcription Factors: chemistry
650 _ 2 |2 MeSH
|a Transcription Factors: genetics
650 _ 2 |2 MeSH
|a Transcription Factors: metabolism
650 _ 7 |0 0
|2 NLM Chemicals
|a Amino Acids, Basic
650 _ 7 |0 0
|2 NLM Chemicals
|a Bacterial Proteins
650 _ 7 |0 0
|2 NLM Chemicals
|a CitB protein, Klebsiella pneumoniae
650 _ 7 |0 0
|2 NLM Chemicals
|a DNA, Bacterial
650 _ 7 |0 0
|2 NLM Chemicals
|a Escherichia coli Proteins
650 _ 7 |0 0
|2 NLM Chemicals
|a Ligands
650 _ 7 |0 0
|2 NLM Chemicals
|a Transcription Factors
650 _ 7 |0 77-92-9
|2 NLM Chemicals
|a Citric Acid
650 _ 7 |0 EC 2.7.-
|2 NLM Chemicals
|a Protein Kinases
650 _ 7 |0 EC 2.7.1.-
|2 NLM Chemicals
|a dpiB protein, E coli
650 _ 7 |a J
|2 WoSType
700 1 _ |a Reinelt, S.
|b 1
|0 P:(DE-HGF)0
700 1 _ |a Kaspar, S.
|b 2
|u FZJ
|0 P:(DE-Juel1)VDB723
700 1 _ |a Scapozza, L.
|b 3
|0 P:(DE-HGF)0
700 1 _ |a Bott, M.
|b 4
|u FZJ
|0 P:(DE-Juel1)128943
773 _ _ |a 10.1021/bi0340595
|g Vol. 42, p. 5917 - 5924
|p 5917 - 5924
|q 42<5917 - 5924
|0 PERI:(DE-600)1472258-6
|t Biochemistry
|v 42
|y 2003
|x 0006-2960
856 7 _ |u http://dx.doi.org/10.1021/bi0340595
|u http://hdl.handle.net/2128/2417
856 4 _ |u https://juser.fz-juelich.de/record/30967/files/30204.pdf
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