Tris Buffer Reactivity with Low-Molecular-Weight Aldehydes: NMR Characterization of the Reactions of Glyceraldehyde-3-Phosphate

Bubb, William A.; Berthon, Hilary A.; Kuchel, Philip W.

doi:10.1006/bioo.1995.1010

Cited by 34 publications

(21 citation statements)

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“…In Tris buffer, K m D-GAP values increased with increasing Tris concentration. This can be attributed to D,L-GAP instability in Tris, presumably a consequence of the reactivity of the aldehyde group of D,L-GAP toward the primary amine group of Tris (36). Citrate buffer was found to inhibit DXP synthase and IspC (supplemental Fig.…”

Section: Discussionmentioning

confidence: 99%

1-Deoxy-d-xylulose 5-Phosphate Synthase Catalyzes a Novel Random Sequential Mechanism

Brammer

Smith

Wade

et al. 2011

Journal of Biological Chemistry

138

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Background: 1-Deoxy-D-xylulose 5-phosphate (DXP) synthase is a thiamine diphosphate (ThDP)-dependent enzyme in pathogen isoprenoid biosynthesis and a potential drug target. Results: Tryptophan fluorescence and kinetic analyses show that donor and acceptor substrates bind reversibly and independently to DXP synthase. Conclusion: DXP synthase catalyzes a novel, ThDP-dependent, random sequential mechanism. Significance: Targeting the unique kinetic mechanism of DXP synthase could lead to new anti-infective agents.

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Section: Discussionmentioning

confidence: 99%

1-Deoxy-d-xylulose 5-Phosphate Synthase Catalyzes a Novel Random Sequential Mechanism

Brammer

Smith

Wade

et al. 2011

Journal of Biological Chemistry

138

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“…To fulfill its function, an ideal biological buffer should not only possess the correct pK a and buffer capacity but should also produce no adverse effects on biochemical reactions [1][2][3], and there may be no one ideal buffer for a given pH range. For example, TRIS [1], HEPES, and Tricine buffers have been shown to react with radicals [4][5][6][7], and TRIS has also been shown to react with aldehydes [8][9][10][11][12]. It is important to carry out measurements in more than one kind of buffer to establish which buffers cause least disturbance to the system under study, the usual criterion being that the observed reaction rates or transformations are maximal.…”

Section: Introductionmentioning

confidence: 99%

Buffer interactions: Densities and solubilities of some selected biological buffers in water and in aqueous 1,4-dioxane solutions

Taha

Lee

2009

Biochemical Engineering Journal

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“…In the experiments reported here the cells were suspended in physiological saline. We have observed that Tris buffer reacts with RBC metabolites (19) and it is conceivable that a combination of the centrifugation conditions and the buffer contribute to the volume change reported previously (8).…”

Section: Discussionmentioning

confidence: 68%

³¹P MAS‐NMR of human erythrocytes: Independence of cell volume from angular velocity

Kuchel

Bubb

Ramadan

et al. 2004

Magnetic Resonance in Med

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4 31P magic angle spinning NMR (MAS-NMR) spectra were obtained from suspensions of human red blood cells (RBCs) that contained the cell-volume-sensitive probe molecule, dimethyl methylphosphonate (DMMP). A mathematical representation of the spectral-peak shape, including the separation and widthat-half-height in the 31 P NMR spectra, as a function of rotor speed, enabled us to explore the extent to which a change in cell volume would be reflected in the spectra if it occurred. We concluded that a fractional volume change in excess of 3% would have been detected by our experiments. Thus, the experiments indicated that the mean cell volume did not change by this amount even at the highest spinning rate of 7 kHz. The mean cell volume and intracellular Key words: 31 P MAS-NMR; magic angle spinning; red blood cells; dimethyl methylphosphonateMagic angle spinning NMR (MAS-NMR) spectra of intact tissue samples have the potential to yield high-resolution "fingerprints" of component metabolites (1-3), while avoiding the risk of decomposition of metabolites that may be associated with some extraction procedures. It has been suggested that MAS conditions may cause tissue damage (2,3), but this problem could in principle be eliminated with slow spinning (4 -6). However, the implementation of these methods is neither straightforward nor even accessible on all MAS spectrometers, and it is not known whether cells will behave in metabolically normal ways under even moderate spin rates.In their observation that MAS NMR could be employed to resolve intra-and extracellular water signals in human erythrocytes (red blood cells; RBCs), Humpfer et al. (7) found no evidence of damage to the cells due to the centrifugal forces in the rotor. Moreover, it was noted that the preparation of RBCs usually involves centrifugal forces comparable to those encountered in the MAS conditions that were employed. Yet Yedgar and co-workers reported (8,9) a number of changes in the physical properties of RBCs that had been subjected to centrifugation conditions that theoretically yield similar forces to those encountered in MAS rotors. Furthermore, there have been reports (e.g., 10,11) that, even under static conditions the NMR lineshapes from RBC suspensions are susceptible to cell settling.In an attempt to understand and interpret these apparent contradictions we studied the effect on cell volume of human RBCs in suspensions under 31 P NMR MAS conditions at 162 MHz. We added dimethyl methylphosphonate (DMMP) to the cells to monitor their mean volume (12,13) during MAS at 250 Hz to 7 kHz. This ability to monitor cell volume during exposure to the centrifugal forces associated with the spinning process is a unique attribute of this particular NMR experiment.While it is known that the 1 H NMR resonance of water in an RBC suspension is split into two peaks (7), the effect was only recently identified as being due to both a difference in the magnetic susceptibility and a difference in isotropic chemical shift due to the different average extent of hydrog...

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Tris Buffer Reactivity with Low-Molecular-Weight Aldehydes: NMR Characterization of the Reactions of Glyceraldehyde-3-Phosphate

Cited by 34 publications

References 0 publications

1-Deoxy-d-xylulose 5-Phosphate Synthase Catalyzes a Novel Random Sequential Mechanism

1-Deoxy-d-xylulose 5-Phosphate Synthase Catalyzes a Novel Random Sequential Mechanism

Buffer interactions: Densities and solubilities of some selected biological buffers in water and in aqueous 1,4-dioxane solutions

³¹P MAS‐NMR of human erythrocytes: Independence of cell volume from angular velocity

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Tris Buffer Reactivity with Low-Molecular-Weight Aldehydes: NMR Characterization of the Reactions of Glyceraldehyde-3-Phosphate

Cited by 34 publications

References 0 publications

1-Deoxy-d-xylulose 5-Phosphate Synthase Catalyzes a Novel Random Sequential Mechanism

1-Deoxy-d-xylulose 5-Phosphate Synthase Catalyzes a Novel Random Sequential Mechanism

Buffer interactions: Densities and solubilities of some selected biological buffers in water and in aqueous 1,4-dioxane solutions

31P MAS‐NMR of human erythrocytes: Independence of cell volume from angular velocity

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³¹P MAS‐NMR of human erythrocytes: Independence of cell volume from angular velocity