Structure and Dynamics of the Modular Halves of Escherichia coli Cyclic AMP Receptor Protein

Citations Over Time

Abstract

E. coli cyclic AMP receptor protein, CRP, is a modular protein that consists of a covalent linkage of two common structural domains. To probe the mechanism for intramolecular communications and to define the unique properties acquired by covalent linkage, the structural, and functional properties of the cAMP- and DNA-binding domains of CRP were studied separately as two independent polypeptides. The N-terminal cAMP-binding domain (alpha-CRP), including S-CRP and CH-CRP, which were generated by digestion of CRP by subtilisin and chymotrypsin, respectively, are mainly populated by beta-sheets. The C-terminal DNA-binding domain, designated as beta-CRP, consists of mostly alpha-helices. The residues of S-CRP and CH-CRP are from 1 to 116 and 1 to 136 of intact wild-type CRP, and those of beta-CRP are from 108 to 209. The secondary structures of alpha-CRP and beta-CRP were monitored by FT-IR, and they are similar to those of the corresponding parts in intact wild-type CRP. Results from hydrogen-deuterium exchange experiments indicated that beta-CRP is more dynamic than alpha-CRP. In an earlier study, it was shown that alpha-CRP retains the function of binding cAMP [Heyduk, E., et al. (1992) Biochemistry 31, 3682-3688]. beta-CRP was able to bind to DNA, although only weakly, and was not sequence specific. Thus, a covalent linkage between the two domains is essential for the realization of the intramolecular signal transmission between the domains triggered by ligand binding. The acquisition of this unique property is intimately associated with the dynamics of the molecule.

Related Papers

• → ChemInform Abstract: Synthesis of Fused Heterocycles via Preferential Hydroamination of O‐Haloarylalkynes over N‐Arylation and Successive Intramolecular C—C Bond Formation.(2015)1 cited
• → ChemInform Abstract: Intramolecular Ring‐Opening of Cyclopropanones by Enolate Anions.(1995)
• → ChemInform Abstract: Transition Metal Promoted Intramolecular Cyclizations of α,α‐Dichloro Esters and Acids.(1987)
• → ChemInform Abstract: Direct Formation of a Substituted (5.5.5.5)Fenestrane by Intramolecular Arene‐Olefin Photocycloaddition.(1989)
• → CuI‐Catalyzed Intramolecular O‐Vinylation of Carbonyl Compounds.(2005)