Related ArticlesRelation of enzyme activity to local/global stability of murine adenosine deaminase: 19F NMR studies.
J Mol Biol. 2005 Jan 21;345(3):599-610
Authors: Shu Q, Frieden C
Adenosine deaminase (ADA, EC 3.5.4.4) is a ubiquitous (beta/alpha)8-barrel enzyme crucial for purine metabolism and normal immune competence. In this study, it was observed that loss of enzyme activity of murine ADA (mADA) precedes the global secondary and tertiary structure transition when the protein is exposed to denaturant. The structural mechanism for this phenomenon was probed using site-specific 19F NMR spectroscopy in combination with [6-19F]tryptophan labeling and inhibitor binding. There are four tryptophan residues in mADA and all are located more than 12 A from the catalytic site. The 19F NMR spectra of [6-19F]Trp-labelled mADA show that the urea-induced chemical shift change of 19F resonance of W161, one of the four tryptophan 19F nuclei, correlates with the loss of enzyme activity. The urea-induced chemical shift change of another 19F resonance of W117 correlates with the change of the apparent rate constant for the binding of transition-state analogue inhibitor deoxycoformycin to the enzyme. On the other hand, the chemical environment of the local region around W264 does not change significantly, as a consequence of perturbation by low concentrations of urea or substrate analog. The results indicate that different regions of mADA have different local stability, which controls the activity and stability of the enzyme. The results provide new insights into the relationship between the function of a protein and its conformational flexibility as well as its global stability. This study illustrates the advantage of 19F NMR spectroscopy in probing site-related conformational change information in ligand binding, enzymatic activity and protein folding.
[Question from NMRWiki Q&A forum] relation of T2 relaxation time to gradient strengths
relation of T2 relaxation time to gradient strengths
to detect short T2 NMR relaxation time, we use high gradient strengths. but how? what is the relation? what is the mechanism behind it? I am interested more in physical picture rather than mathematical description.
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10-21-2011 09:52 AM
NMR solution structure of human VRK1 reveals the C-terminal tail essential for structural stability and autocatalytic activity.
NMR solution structure of human VRK1 reveals the C-terminal tail essential for structural stability and autocatalytic activity.
NMR solution structure of human VRK1 reveals the C-terminal tail essential for structural stability and autocatalytic activity.
J Biol Chem. 2011 May 3;
Authors: Shin J, Chakraborty G, Bharatham N, Kang C, Tochio N, Koshiba S, Kigawa T, Kim W, Kim KT, Yoon HS
Vaccinia-related kinase 1 (VRK1) is one of the mitotic kinases which play important roles in cell cycle, nuclear condensation and transcription regulation. Kinase...
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05-06-2011 12:02 PM
[NMR paper] Urea-dependent unfolding of murine adenosine deaminase: sequential destabilization as
Urea-dependent unfolding of murine adenosine deaminase: sequential destabilization as measured by 19F NMR.
Related Articles Urea-dependent unfolding of murine adenosine deaminase: sequential destabilization as measured by 19F NMR.
Biochemistry. 2004 Feb 17;43(6):1432-9
Authors: Shu Q, Frieden C
Murine adenosine deaminase (mADA) is a 40 kDa (beta/alpha)(8)-barrel protein consisting of eight central beta-strands and eight peripheral alpha-helices containing four tryptophan residues. In this study, we investigated the urea-dependent behavior of...
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11-24-2010 09:25 PM
[NMR paper] Cytosine deaminase and thymidine kinase gene therapy in a Dunning rat prostate tumour
Cytosine deaminase and thymidine kinase gene therapy in a Dunning rat prostate tumour model: absence of bystander effects and characterisation of 5-fluorocytosine metabolism with 19F-NMR spectroscopy.
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Gene Ther. 2002 Dec;9(23):1564-75
Authors: Corban-Wilhelm H, Hull WE, Becker G, Bauder-Wüst U, Greulich D, Debus J
The rat...
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11-24-2010 08:58 PM
[NMR paper] Local fluctuations and global unfolding of partially folded BPTI detected by NMR.
Local fluctuations and global unfolding of partially folded BPTI detected by NMR.
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Biophys Chem. 1997 Feb 28;64(1-3):45-57
Authors: Barbar E, LiCata VJ, Barany G, Woodward C
The protein Abu is a chemically synthesized variant of bovine pancreatic trypsin inhibitor (BPTI) with the 14-38 disulfide bond intact and cysteines 5, 30, 51, and 55 replaced by alpha-amino-n-butyric acid (Abu). At 1-6 degrees C and pH 4.5-6.5, Abu is partially folded with a...
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08-22-2010 03:31 PM
[NMR paper] Local fluctuations and global unfolding of partially folded BPTI detected by NMR.
Local fluctuations and global unfolding of partially folded BPTI detected by NMR.
Related Articles Local fluctuations and global unfolding of partially folded BPTI detected by NMR.
Biophys Chem. 1997 Feb 28;64(1-3):45-57
Authors: Barbar E, LiCata VJ, Barany G, Woodward C
The protein Abu is a chemically synthesized variant of bovine pancreatic trypsin inhibitor (BPTI) with the 14-38 disulfide bond intact and cysteines 5, 30, 51, and 55 replaced by alpha-amino-n-butyric acid (Abu). At 1-6 degrees C and pH 4.5-6.5, Abu is partially folded with a...
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08-22-2010 03:03 PM
[NMR paper] NMR studies of the solution properties of recombinant murine interleukin-6.
NMR studies of the solution properties of recombinant murine interleukin-6.
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Biochim Biophys Acta. 1995 Jun 12;1249(2):189-203
Authors: Morton CJ, Bai H, Zhang JG, Hammacher A, Norton RS, Simpson RJ, Mabbutt BC
The effects of solvent, pH and temperature on the 1H-NMR spectra of recombinant murine interleukin-6 (IL-6) are described. Assignments made from two-dimensional homonuclear spectra are presented for resonances of the fifteen aromatic amino-acid...
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08-22-2010 03:41 AM
[NMR paper] Solution structure of murine epidermal growth factor determined by NMR spectroscopy a
Solution structure of murine epidermal growth factor determined by NMR spectroscopy and refined by energy minimization with restraints.
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Biochemistry. 1992 Jan 14;31(1):236-49
Authors: Montelione GT, Wüthrich K, Burgess AW, Nice EC, Wagner G, Gibson KD, Scheraga HA
The solution structure of murine epidermal growth factor (mEGF) at pH 3.1 and a temperature of 28 degrees C has been determined...
Relation of enzyme activity to local/global stability of murine adenosine deaminase:
Linear Mode
