Conformational selection of <i>syn</i>‐cAMP upon binding to the cAMP receptor protein

Gronenborn, Angela M.; Clore, G. Marius; Blazy, B.; Baudras, Alain

doi:10.1016/0014-5793(81)81237-9

Cited by 31 publications

(12 citation statements)

References 14 publications

(5 reference statements)

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“…Mutational analysis confirms the importance of side chains interacting with cAMP in the structures (9)(10)(11)(12)(13). In contrast to the crystallographic observations, transfer nuclear Overhauser effect (NOE) NMR measurements showed a rapidly exchanging cAMP molecule bound to CAP in the syn conformation (14,15).…”

mentioning

confidence: 54%

The structure of a CAP–DNA complex having two cAMP molecules bound to each monomer

Passner

Steitz

1997

Proc. Natl. Acad. Sci. U.S.A.

173

167

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The 2.2 A resolution crystal structure of the Escherichia coli catabolite gene activator protein (CAP) complexed with cAMP and a 46-bp DNA fragment reveals a second cAMP molecule bound to each protein monomer. The second cAMP is in the syn conformation and is located on the DNA binding domain interacting with the helix-turn-helix, a beta-hairpin from the regulatory domain and the DNA (via water molecules). The presence of this second cAMP site resolves the apparent discrepancy between the NMR and x-ray data on the conformation of cAMP, and explains the cAMP concentration-dependent behaviors of the protein. In addition, this site's close proximity to mutations affecting transcriptional activation and its water-mediated interactions with a DNA recognition residue (E181) and DNA raise the possibility that this site has biological relevance.

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mentioning

confidence: 54%

The structure of a CAP–DNA complex having two cAMP molecules bound to each monomer

Passner

Steitz

1997

Proc. Natl. Acad. Sci. U.S.A.

173

167

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“…Anderson et al (2) have presented evidence that the 2'-hydroxyl and cyclic 3',5'-phosphoryl groups are important determinants in the binding of a nucleotide to wild-type CRP, whereas the N6-amino group of cAMP plays a role in promoting the structural alteration that activates the protein as a positive control element. Gronenborn et al (14) have recently implicated an interaction between wild-type CRP and the adenine of cAMP in the conformational selection of syn-cAMP upon the binding of the nucleotide to the protein. Neither cGMP nor cCMP has a functional group comparable to the N6-amino group of cAMP and, unlike cAMP, the nucleotides are bound by both wild-type and mutant CRP with similar affinities.…”

Section: Resultsmentioning

confidence: 99%

Mechanism of CRP-mediated cya suppression in Escherichia coli

Harman¹,

Dobrogosz²

1983

J Bacteriol

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Escherichia coli strain NCR30 contains a cya lesion and a second-site cya suppressor mutation that lies in the crp gene. NCR30 shows a pleiotropic phenotypic reversion to the wild-type state in expressing many operons that require the cyclic AMP (cAMP)-cAMP receptor protein (CRP) complex for positive control. In vivo 3-galactosidase synthesis in NCR30 was sensitive to glucose-mediated repression, which was relieved not only by cAMP but also by cyclic GMP and cyclic CMP. The CRP isolated from NCR30 differed from the protein isolated from wild-type E. coli in many respects. The mutant protein bound cAMP with four to five times greater affinity than wild-type CRP. Protease digestion studies indicated that native NCR30 CRP exists in the cAMP-CRP complex-like conformation. The protein conferred a degree of cAMP independence on the in vitro synthesis of P-galactosidase. In addition, the inherent positive control activity of the mutant protein in vitro was enhanced by those nucleotides that stimulate in vivo P-galactosidase synthesis in NCR30. The results of this study supported the conclusion that the crp allele of NCR30 codes for a protein having altered effector specificity yet capable of promoting positive control over catabolite-sensitive operons in the absence of an effector molecule.

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“…This phenomenon is defined as a process in which the binding of two components (biological macromolecules to small molecular ligands or to each other) occurs in a milieu containing a pre‐existing equilibrium of multiple macromolecular conformations, among which ligands can select the best fitting ones 22. The appearance of the term conformational selection goes back 30 years 23, 24, however it has only been widely used in the last decade since Nussinov and co‐workers 25–28 published several papers on the topic. In the last few years the number of publications in this field has increased exponentially 22, 29–32 (Fig.…”

Section: Introductionmentioning

confidence: 99%