Ligand-induced structural changes to maltodextrin-binding protein as studied by solution NMR spectroscopy.

نویسندگان

  • J Evenäs
  • V Tugarinov
  • N R Skrynnikov
  • N K Goto
  • R Muhandiram
  • L E Kay
چکیده

Solution NMR studies on the physiologically relevant ligand-free and maltotriose-bound states of maltodextrin-binding protein (MBP) are presented. Together with existing data on MBP in complex with beta-cyclodextrin (non-physiological, inactive ligand), these new results provide valuable information on changes in local structure, dynamics and global fold that occur upon ligand binding to this two-domain protein. By measuring a large number of different one-bond residual dipolar couplings, the domain conformations, critical for biological function, were investigated for all three states of MBP. Structural models of the solution conformation of MBP in a number of different forms were generated from the experimental dipolar coupling data and X-ray crystal structures using a quasi-rigid-body domain orientation algorithm implemented in the structure calculation program CNS. Excellent agreement between relative domain orientations in ligand-free and maltotriose-bound solution conformations and the corresponding crystal structures is observed. These results are in contrast to those obtained for the MBP/beta-cyclodextrin complex where the solution state is found to be approximately 10 degrees more closed than the crystalline state. The present study highlights the utility of residual dipolar couplings for orienting protein domains or macromolecules with respect to each other.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Orienting domains in proteins using dipolar couplings measured by liquid-state NMR: differences in solution and crystal forms of maltodextrin binding protein loaded with beta-cyclodextrin.

Protein function is often regulated by conformational changes that occur in response to ligand binding or covalent modification such as phosphorylation. In many multidomain proteins these conformational changes involve reorientation of domains within the protein. Although X-ray crystallography can be used to determine the relative orientation of domains, the crystal-state conformation can refle...

متن کامل

Synthesis, Characterization and interaction Studies of 1-(3-bromophenyl azo) 2,7-dihydroxy naphthalene, (BPADHN) with calf thymus deoxy ribo nucleic acid (ct-DNA)

In this study at first , an azo dye, 2,7- naphthalenediol, 2-[(4-Bromophenyl)azo (BPAND) as a ligand has been synthesized by addition of p-Bromoaniline to the modified montomorillonite K10 clay. This ligand was characterized using 1H-NMR, UV-Vis and IR spectroscopies. Subsequently, its interaction with calf thymus deoxyribonucleicacid ,ct-DNA was investigated in 5 mM phosphate buffer solution, ...

متن کامل

SPECTROSCOPIC EVALUATION OF THE INTERACTION OF A TETRAZOLE DERIVATIVE SYNTHESIZED BY SEMI-GREEN METHOD WITH CALF THYMUS DNA AND BOVINE SERUM PROTEIN

Background & Aims: In recent decades, the application of tetrazole structures in various fields of medicine and industry has become very important, because they can cause structural and thus functional changes in the proteins. In this article, the effect of a new tetrazole derivative on calf thymus DNA (Ct-DNA) as well as on bovine serum albumin protein (BSA) in the solution was determined usin...

متن کامل

Label-free quantitative 1H NMR spectroscopy to study low-affinity ligand–protein interactions in solution: A contribution to the mechanism of polyphenol-mediated astringency

Nuclear magnetic resonance (NMR) spectroscopy is well-established in assessing the binding affinity between low molecular weight ligands and proteins. However, conventional NMR-based binding assays are often limited to small proteins of high purity and may require elaborate isotopic labeling of one of the potential binding partners. As protein-polyphenol complexation is assumed to be a key even...

متن کامل

Thermal aggregation of egg white proteins as affected by saccharides

Thermal characteristics of egg white proteins (EWP) may differ in the presence of saccharides. Therefore, the influence of saccharides including carboxymethyl cellulose (CMC), pectin, sucrose and maltodextrin and heating time on physicochemical characteristics of EWP as a whole were studied. Investigation of Heat Coagulation Time (HCT), solubility, turbidity and protein secondary structure of h...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • Journal of molecular biology

دوره 309 4  شماره 

صفحات  -

تاریخ انتشار 2001