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129Xe 'r2 relaxation switch type' magnetic resonance molecular probe for recognition of lysine decarboxylase

A technology of lysine decarboxylase and molecular probe, applied in the field of 129Xe "r2 relaxation efficiency switch type" magnetic resonance molecular probe, achieving high-sensitivity recognition and strong penetrability

Active Publication Date: 2019-05-14
WUHAN INST OF PHYSICS & MATHEMATICS CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The determination method is simple and the determination result is accurate, but it needs to use the toxic organic solvent toluene

Method used

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  • 129Xe 'r2 relaxation switch type' magnetic resonance molecular probe for recognition of lysine decarboxylase
  • 129Xe 'r2 relaxation switch type' magnetic resonance molecular probe for recognition of lysine decarboxylase
  • 129Xe 'r2 relaxation switch type' magnetic resonance molecular probe for recognition of lysine decarboxylase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1 measures the relaxation efficiency of ammonium acetate buffer

[0036] (1) Get ammonium acetate and dissolve it in ultrapure water, and configure the solutions whose concentration of ammonium acetate is 0mM, 2mM, 4mM, 6mM, 8mM, and 10mM to be respectively recorded as solution 1, solution 2, solution 3, solution 4, solution 5, Solution 6;

[0037] (2) Each 2 mL of solution 1, solution 2, solution 3, solution 4, solution 5 and solution 6 prepared in the present embodiment was obtained, and the 400MHz magnetic resonance spectrometer was used to 129 In Xe spectrum mode, it is used in conjunction with a hyperpolarized device (the gas heating temperature is 135°C, the gas flow rate is 0.12 standard liters / minute, and the gas composition is 2% Xe, 8% N in volume percentage 2 and 90% He, of which Xe is 26.4% of natural abundance 129 Xe), the experimental temperature is 298K, the test solution 129 T of Xe in solution 1, solution 2, solution 3, solution 4, solutio...

Embodiment 2

[0038] Embodiment 2 tests the relaxation efficiency of lysine solution

[0039] (1) Get ammonium acetate and be dissolved in ultrapure water, be configured into the solution that ammonium acetate concentration is 10mM, be recorded as solution 1;

[0040] (2) Take 5 parts of lysine and dissolve them in solution 1 respectively, sonicate until the lysine is completely dissolved, and configure the concentration of lysine to be 0.2μM, 0.4μM, 0.6μM, 0.8μM, 1.0μM, and use acetic acid and Ammonia liquor adjusts the pH of solution respectively, makes the pH of solution be 6.0, is recorded as solution 2, solution 3, solution 4, solution 5, solution 6 respectively;

[0041] (3) 2 mL of solution 1, solution 2, solution 3, solution 4, solution 5, and solution 6 prepared in this embodiment are each taken in 2 mL with a 400 MHz magnetic resonance spectrometer 129 In Xe spectrum mode, it is used in conjunction with a hyperpolarized device (the gas heating temperature is 135°C, the gas flow r...

Embodiment 3

[0042] Embodiment 3 measures the relaxation efficiency of lysine decarboxylase solution

[0043] (1) Get ammonium acetate and be dissolved in ultrapure water, be configured into the solution that ammonium acetate concentration is 10mM, be recorded as solution 1;

[0044] (2) Take 4 parts of lysine decarboxylase, dissolve them in solution 1 respectively, configure the concentration of lysine decarboxylase as 0.5μg / mL, 1μg / mL, 1.5μg / mL, 2μg / mL, add acetic acid and ammonia water Adjust the pH of the solution respectively, so that the pH of the solution is 6.0, which are respectively recorded as solution 2, solution 3, solution 4, and solution 5;

[0045] (3) Each 2 mL of solution 1, solution 2, solution 3, solution 4 and solution 5 prepared in the present embodiment was obtained, and the 400MHz magnetic resonance spectrometer was used to 129 In Xe spectrum mode, it is used in conjunction with a hyperpolarized device (the gas heating temperature is 135°C, the gas flow rate is 0.1...

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Abstract

The invention belongs to the technical field of biochemical analysis, and particularly relates to a 129Xe 'r2 relaxation efficiency switch type' magnetic resonance molecular probe for recognition of lysine decarboxylase. The probe includes cucurbit [6] urea and lysine. The principle of the molecular probe is as follows: a weak gourd [6] urea-binding substrate (lysine) is converted by lysine decarboxylase into a strong gourd [6] urea-binding product (cadaverine). Before an enzymatic reaction, the relaxation efficiency of the molecular probe is relatively high due to the chemical exchange of 129Xe in cucurbita [6] urea and 129Xe in a soluble state. After lysine decarboxylase is added, lysine is gradually converted into cadaverine by lysine decarboxylase, and the cucurbita [6] urea is occupied by cadaverine. Finally, the relaxation efficiency of the molecular probe becomes almost zero. Recognition of lysine decarboxylase is achieved by changing the relaxation efficiency of molecular probes.

Description

technical field [0001] The invention relates to the technical field of biochemical analysis, in particular to a method for recognizing lysine decarboxylase 129 Xe"r 2 Relaxation Efficiency Switching" Magnetic Resonance Molecular Probes. Background technique [0002] Lysine decarboxylase can catalyze the decarboxylation of L-lysine to generate cadaverine (also known as "1,5-pentanediamine") and carbon dioxide, and is a key enzyme in the biological cadaverine synthesis pathway. The cadaverine produced by its decarboxylation reacts with oxygen free radicals to help eliminate the damage of free radicals in organisms to the body, thereby protecting microbial cells from oxygen poisoning. (Igarashi K, Kashiwagi K. Modulation of cellular function by polyamines. Int J Biochem Cell Biol. 2010, 42(1):39-51.). Endogenous decarboxylation by lysine decarboxylase and accumulation of high intracellular concentrations of pentamethylenediamine can lead to decreased outer membrane permeabil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/527G01N24/08
Inventor 周欣张斌郭茜旎孙献平刘买利叶朝辉
Owner WUHAN INST OF PHYSICS & MATHEMATICS CHINESE ACADEMY OF SCI
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