title: Binding modes of cyclic AMP and environments of tryptophan residues in 1:1 and 1:2 complexes of cyclic AMP receptor protein and cyclic AMP.
abstract: Cyclic AMP (cAMP) receptor protein (CRP) forms 1:1 and 1:2 complexes with cAMP, and the former complex is considered to be the most active form of CRP in binding to specific DNA sequences and in modulating gene transcription by RNA polymerases. We examine the cAMP binding modes and structural changes of CRP upon cAMP binding by UV resonance Raman spectroscopy. The Raman spectra of CRP-(cAMP)(1) and CRP-(cAMP)(2) extracted from those of CRP-cAMP mixtures at varied mixing ratios clearly show that the hydrogen bonding state and the conformation of cAMP in both complexes in solution are very similar to those found in the X-ray crystal structure of CRP-(cAMP)(2), which is evidence that the cAMP binding mode does not differ between the two complexes. The environmental hydrophobicity of Trp85 monitored by UV resonance Raman intensity shows a significant decrease upon binding of the first cAMP molecule, whereas no further change occurs in the second cAMP binding step. The environmental change of Trp85 suggests an opening of the cleft between the N-terminal cAMP and C-terminal DNA binding domains in the process of CRP activation by binding of a single cAMP molecule.
authors: Graduate School of Pharmaceutical Sciences, Tohoku University, Aobayama, Sendai 980-8578, Japan.
authors: Naoko
authors: N
authors: Fujimoto
authors: Akira
authors: A
authors: Toyama
authors: Hideo
authors: H
authors: Takeuchi
mesh_terms: D000242:Cyclic AMP / Q000737:chemistry* / Q000378:metabolism*; D002373:Cyclic AMP Receptor Protein / Q000737:chemistry*; D004926:Escherichia coli / Q000378:metabolism; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D013059:Spectrum Analysis, Raman; D014364:Tryptophan / Q000737:chemistry*; D014466:Ultraviolet Rays
chemical_list: D002373:Cyclic AMP Receptor Protein; D014364:Tryptophan; D000242:Cyclic AMP