Microcalorimetric study of magnesium-adenosine triphosphate ternary complex

Sari, J.C.; Hadida, Michèle; Chauvet-Monges, Anne-Marie; Crevat, A.

doi:10.1007/bf00745018

Cited by 14 publications

(5 citation statements)

References 27 publications

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“…The charge balance is maintained because the elongated RNA molecule gains one more negative charge with each nucleotide addition. The association constants for magnesium to NTPs (Sari et al, 1982) and to PP i (Irani, 1961) are such that these components can be assumed to have one Mg 2+ bound under typical transcription conditions. The use of inorganic pyrophosphatase (PPase) has been shown to be beneficial for obtaining high yields of RNA, presumably because accumulation of PP i byproduct results in inhibition of RNA transcription (Cunningham and Ofengand, 1990).…”

Section: Theory: Transcription Solution Equilibrium Analysismentioning

confidence: 99%

“…where T is the absolute temperature, R is the ideal gas constant, and ∆Hi is the enthalpy of ionization. The numbers in the last column correspond to the following references: (1) Smith and Alberty (1956a), (2) Sari et al (1982), (3) Smith and Alberty (1956b), (4) Irani (1961), (5) Roig et al (1993), (6) Arnold et al (1985), and (7) Gold and Powell (1976…”

Section: Theory: Transcription Solution Equilibrium Analysismentioning

confidence: 99%

“…During transcription, each nucleotide addition results in the formation of an inorganic pyrophosphate ion (PP i ≡ P 2 O 7 4 − ) and the release of a proton: The charge balance is maintained because the elongated RNA molecule gains one more negative charge with each nucleotide addition. The association constants for magnesium to NTPs (Sari et al, 1982) and to PP i (Irani, 1961) are such that these components can be assumed to have one Mg 2+ bound under typical transcription conditions.…”

Section: Theory: Transcription Solution Equilibrium Analysismentioning

confidence: 99%

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Application of Solution Equilibrium Analysis to in Vitro RNA Transcription

Kern

Davis

1997

Biotechnol. Prog.

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Solution equilibrium analysis of in vitro RNA transcription has been applied to examine changes in pH, free magnesium concentration, and concentrations of all chemical ionization species as a transcription reaction proceeds. With this method, the progress of a transcription reaction can be accurately determined as a function of measured pH. In addition, it is demonstrated that this method has significant value as a tool for achieving improved understanding of the effects of varying solution conditions on the dynamics of RNA transcription. Magnesium concentration was found to be a critical factor for efficient transcription. Below 5 mM free Mg2+ concentration, the transcription rate and the efficiency at which nucleoside triphosphates (NTPs) are incorporated are greatly reduced. While inorganic pyrophosphate (PPi), a byproduct of the reaction, was found to directly inhibit the rate of transcription, its detrimental effects on transcription were determined to be primarily due to sequestering of magnesium. The PPi forms a precipitate with magnesium which was determined to have a molar composition of 2:1 of Mg:PPi. Transcription rate and efficiency of NTP incorporation are also reduced with increasing ionic strength. It is shown that these reductions can be partially alleviated by replacing chloride with acetate anions.

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Section: Theory: Transcription Solution Equilibrium Analysismentioning

confidence: 99%

Section: Theory: Transcription Solution Equilibrium Analysismentioning

confidence: 99%

Section: Theory: Transcription Solution Equilibrium Analysismentioning

confidence: 99%

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Application of Solution Equilibrium Analysis to in Vitro RNA Transcription

Kern

Davis

1997

Biotechnol. Prog.

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“…We also include Mg 2+ (aq) , or more specifically, [Mg(H 2 O) 6 ] 2+ (aq) , the octahedral complex configuration adopted by the free ion in aqueous solution . This ion interacts with one or more phosphate units to form various Mg 2+ –ATP (aq) complexes, − such as [Mg(ATP) 2 ] 6– (aq) , ,− Mg 2 ATP (aq) , and [Mg 2 (ATP) 2 ] 4– (aq) . , For [MgATP] 2– (aq) , a closed form has also been proposed, where the metal ion interacts not only with O atoms from the phosphate units but also, presumably through a water molecule, with a N atom from the adenine unit. The “closed form”/“open form” ratio was reported to be 1:10 in aqueous solution. ,− A schematic representation of this configuration is shown in Figure S1 in the Supporting Information (SI).…”

Section: Introductionmentioning

confidence: 99%

“… 23 This ion interacts with one or more phosphate units to form various Mg 2+ –ATP (aq) complexes, 24 − 26 such as [Mg(ATP) 2 ] 6– (aq) , 24 , 27 − 30 Mg 2 ATP (aq) , 31 and [Mg 2 (ATP) 2 ] 4– (aq) . 24 , 32 For [MgATP] 2– (aq) , a closed form has also been proposed, where the metal ion interacts not only with O atoms from the phosphate units but also, presumably through a water molecule, with a N atom from the adenine unit. The “closed form”/“open form” ratio was reported to be 1:10 in aqueous solution.…”

Section: Introductionmentioning

confidence: 99%

How Does Mg²⁺_(aq) Interact with ATP_(aq)? Biomolecular Structure through the Lens of Liquid-Jet Photoemission Spectroscopy

Mudryk,

Lee,

Tomaník

et al. 2024

J. Am. Chem. Soc.

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Liquid-jet photoemission spectroscopy (LJ-PES) allows for a direct probing of electronic structure in aqueous solutions. We show the applicability of the approach to biomolecules in a complex environment, exploring site-specific information on the interaction of adenosine triphosphate in the aqueous phase (ATP (aq) ) with magnesium (Mg 2+ (aq) ), highlighting the synergy brought about by the simultaneous analysis of different regions in the photoelectron spectrum. In particular, we demonstrate intermolecular Coulombic decay (ICD) spectroscopy as a new and powerful addition to the arsenal of techniques for biomolecular structure investigation. We apply LJ-PES assisted by electronic-structure calculations to study ATP (aq) solutions with and without dissolved Mg 2+ . Valence photoelectron data reveal spectral changes in the phosphate and adenine features of ATP (aq) due to interactions with the divalent cation. Chemical shifts in Mg 2p, Mg 2s, P 2p, and P 2s core-level spectra as a function of the Mg 2+ /ATP concentration ratio are correlated to the formation of [Mg(ATP) 2 ] 6− (aq) , [MgATP] 2− (aq) , and [Mg 2 ATP] (aq) complexes, demonstrating the element sensitivity of the technique to Mg 2+ −phosphate interactions. The most direct probe of the intermolecular interactions between ATP (aq) and Mg 2+ (aq) is delivered by the emerging ICD electrons following ionization of Mg 1s electrons. ICD spectra are shown to sensitively probe ligand exchange in the Mg 2+ −ATP (aq) coordination environment. In addition, we report and compare P 2s data from ATP (aq) and adenosine mono-and diphosphate (AMP (aq) and ADP (aq) , respectively) solutions, probing the electronic structure of the phosphate chain and the local environment of individual phosphate units in ATP (aq) . Our results provide a comprehensive view of the electronic structure of ATP (aq) and Mg 2+ −ATP (aq) complexes relevant to phosphorylation and dephosphorylation reactions that are central to bioenergetics in living organisms.

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