Nuclear Magnetic Resonance Study of Complexes between CRP and Its 22- and 28-Base-Pair Operators

Lee, Bong Jin; Lee, Tae Woo; Park, Sang Ho; Aiba, Hiroji; Kojima, Chojiro; Kyogoku, Yoshimasa

doi:10.1093/jb/117.2.239

Cited by 28 publications

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“…Kinetic Consideration on PC − Lysine Peptide Binding and Its Effect on Electron Transfer. It is of fundamental importance to know how proteins recognize their electron accepting and/or donating partners, and there have been a number of studies on the electron transfer between proteins. − Electron transfers between PC and cyt f or cyt c have been studied extensively, ,− ,,,,, where the positively charged cyt c has been used as a model for cyt f . Redox reactions between PC and small molecules have also been investigated extensively, and Sykes et al have previously discovered in an elegant way that small inorganic compounds can inhibit the electron transfer between PC and cyt c or cyt f . ,,− …”

Section: Discussionmentioning

confidence: 99%

“…Preparation of Samples. Silene pratensis (white campion) wild-type and negative patch mutant PC's (M1−M4, Table ) were expressed in Escherichia coli and purified by published methods. , Absorption and EPR spectra of site-directed mutant PC’s were the same as those of wild-type PC, indicating that mutations of amino acid residues at the negative patch do not affect the Cu active site in solution . Moreover, the X-ray crystallographic structure of M4 mutant PC showed good correspondence with that of wild-type PC .…”

Section: Methodsmentioning

confidence: 99%

“…On the other hand, on the basis of the crystal structures of oxidized and reduced plant PC's, − two highly conserved sites of PC have been considered as molecular recognition sites for its redox partners, cyt f , cyt c , and PSI: One site is located at the Cu-coordinating, solvent-accessible histidine (Cu-adjacent hydrophobic patch), and the other site is located at another solvent-accessible site containing acidic residues near a tyrosine residue (Cu-remote negative patch) (Figure ). Early studies on electron transfer from reduced PC to [Co(phen) 3 ] 3+ suggested that both are molecular recognition sites for cyt f , , but computational analyses based on electrostatic forces , and other experimental studies − have indicated that the Cu-remote negative patch is the cyt c / f molecular recognition site, while the electron transfer is believed to occur through the hydrophobic patch of PC . Recently, Kostić et al proposed that PC and cyt c or cyt f bind and react with each other in different configurations resulting from the protein−protein interaction termed as the gating process for electron transfer, − showing possible configurations for the diprotein complex by computer simulation. ,

1 Schematic view of the interaction between PC and tetra-Lys.…”

Section: Introductionmentioning

confidence: 99%

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Plastocyanin−Peptide Interactions. Effects of Lysine Peptides on Protein Structure and Electron-Transfer Character

et al. 1998

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Structural change of plastocyanin (PC) due to the interaction with lysine peptides (Lysptd's) has been studied by absorption, resonance Raman, and electrochemical measurements and by measuring the electron transfer between PC and cytochrome c (cyt c) in the presence of Lysptd. Absorption spectral changes which were observed when Lysptd's up to penta-lysine were added to PC solution have been ascribed by resonance Raman studies to the change in the active site Cu−cysteine geometry upon binding of Lysptd to the PC negative patch. The same spectral changes were observed for the PC−cyt c interaction. Electrochemical measurements showed that the redox potential of PC increases upon Lysptd binding, suggesting that Lysptd's induce a structural change in PC through the copper ligating cysteine residue to make the copper site adapted for facile electron transfer. Lysptd's competitively inhibited the electron transfer from reduced cyt c to oxidized PC, which indicated that they function as models of the PC interacting site of proteins. The effects of Lysptd on electron transfer are explained as competitive inhibition due to neutralization of the PC negative patch by formation of PC·Lysptd complexes. The electron-transfer rate from reduced cyt c to oxidized PC and the inhibiting effect of Lysptd decreased upon decreasing the net charge of the negative patch by mutation. The structural change of PC was also found to decrease significantly with these mutants. The present observations strongly support that the PC negative patch is the dominant cyt c/f molecular recognition site and open up the possibility that charged peptides can be used for studying protein−protein interactions in a systematic way.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

1 Schematic view of the interaction between PC and tetra-Lys.…”

Section: Introductionmentioning

confidence: 99%

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Plastocyanin−Peptide Interactions. Effects of Lysine Peptides on Protein Structure and Electron-Transfer Character

et al. 1998

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“…On the other hand, the crystal structures of plant oxidized and reduced PC's have been determined, − and two highly conserved sites have been considered as molecular recognition sites for its redox partners, cyt f and PSI: One site is located at the Cu-coordinating, solvent-accessible histidine (Cu-adjacent hydrophobic patch), and the other site is located at another solvent-accessible site containing acidic residues near a tyrosine residue (Cu-remote negative patch). The negative patch of PC has been indicated to be the cyt f interacting site through electrostatic interaction by recent studies, − whereas electron transfer from PC to P700 is suggested to follow through the hydrophobic patch. , …”

Section: Introductionmentioning

confidence: 99%

Interactions of Cytochrome c and Cytochrome f with Aspartic Acid Peptides

et al. 1999

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Cytochrome c (cyt c) and cytochrome f (cyt f) molecular recognition characters and their structural changes due to complex formation with negatively charged aspartic acid peptides (Aspptd's) have been studied. Changes in the absorption spectrum of cyt c in the Soret region were detected when Aspptd's, up to penta-Asp, were added to the cyt c solution. These changes were the same as those observed when cyt c interacted with plastocyanin (PC), indicating that Aspptd's interacted with cyt c in the same way as PC. Conformational changes of cyt c due to interaction with Aspptd's observed by resonance Raman spectroscopy were similar to those reported for cyt c when bound with its native partner, cytochrome c oxidase. Electrochemical measurements showed that the redox potential of cyt c and cyt f shifted to lower potentials by 7−20 mV upon Aspptd binding, showing the enhancement in the electron donor ability of both cyt c and cyt f upon complex formation with Aspptd. The changes in the absorption spectrum and redox potential increased with the length and concentration of Aspptd. The observed structural and redox changes of cyt c and cyt f are attributed to adduct formations with Aspptd's by electrostatic interactions and suggest that similar changes would occur for cyt c and cyt f when interacting with proteins. Aspptd's, tetra- and penta-aspartic acid, served as competitive inhibitors of the electron transfer from cyt c or cyt f to PC, which was ascribable to the same adduct formation.

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“…CRP is a homo-dimeric protein consisting of 209 amino acids in each monomer (Fig. 1), and functions by binding, in the presence of the allosteric effector cAMP, to specific DNA sites, and interacting with RNA polymerase (2)(3)(4). RNA polymerase is a holoenzyme consisting of a, p, P', and one of several species of CT subunits, and the RNA polymerase a subunit is also a homo-dimeric protein (5,6).…”

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Detection of the Protein-Protein Interaction between Cyclic AMP Receptor Protein and RNA Polymerase, by 13C-Carbonyl NMR

Lee

Won

Park

et al. 2001

Journal of Biochemistry

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Cyclic AMP receptor protein (CRP) plays a key role in the transcription regulation of many prokaryotic genes. Upon the binding of cyclic AMP, CRP is allosterically activated, binds to target DNA sites, and interacts with RNA polymerase. Although the protein-protein interaction between CRP and RNA polymerase is known to be important for the transcription initiation of the target genes, its structural understanding is still lacking, particularly due to the high molecular mass (approximately 120 kDa) of the protein complex. We assigned all of the (13)C-carbonyl resonances of methionine residues in CRP by using the double labeling and the enzyme digestion techniques. The result of (13)C-carbonyl NMR experiment on [(13)C'-Met]-CRP in the presence of both cyclic AMP and RNA polymerase alpha subunit showed that the two proteins interact with each other in solution in the absence of DNA via the region around the residues from Met 157 to Met 163 in CRP. The results also showed the effectiveness of the selective labeling and (13)C-carbonyl NMR spectroscopy in the specific detection of the protein-protein interaction between large molecules.

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Nuclear Magnetic Resonance Study of Complexes between CRP and Its 22- and 28-Base-Pair Operators

Cited by 28 publications

References 0 publications

Plastocyanin−Peptide Interactions. Effects of Lysine Peptides on Protein Structure and Electron-Transfer Character

Plastocyanin−Peptide Interactions. Effects of Lysine Peptides on Protein Structure and Electron-Transfer Character

Interactions of Cytochrome c and Cytochrome f with Aspartic Acid Peptides

Detection of the Protein-Protein Interaction between Cyclic AMP Receptor Protein and RNA Polymerase, by 13C-Carbonyl NMR

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