Compound for virus fluorescence labeling and living imaging and preparation method of compound
A compound and virus technology, applied in palladium organic compounds, platinum group organic compounds, chemical instruments and methods, etc., can solve problems such as not suitable for long-term observation, easy to cause cytotoxicity, and affect the yield of biomolecules
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Embodiment 1
[0041] The preparation of compound shown in embodiment 1, formula I
[0042] refer to figure 1 The shown synthetic route diagram prepares the compound shown in formula I.
[0043] step 1:
[0044]Add 213mg of compound 1 to a 25mL round bottom flask, then dissolve it in 4mL of anhydrous carbon tetrachloride, add 16mg of AIBN (azobisisobutyronitrile) under stirring, then 196mg of NBS (N-bromosuccinimide) was slowly added to the above solution. Then, the temperature of the reaction system was raised to reflux, and the reaction system was monitored by TLC (thin layer chromatography). After the reaction was complete, the reaction system was lowered to room temperature, filtered, the obtained filtrate was rotary evaporated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate=5 / 1) to obtain a white solid 240 mg (Compound 2), the yield was 82%.
[0045] 1 H NMR (400MHz, Chloroform-d) δ7.83–7...
Embodiment 2
[0066] Embodiment 2, the fluorescent labeling of the compound shown in formula I on the polypeptide
[0067] First, the compound is tested on a cysteine-containing polypeptide, and the reaction process is as follows:
[0068] Add 47 μl HO to the Ep tube 2 O, 4 μl 150 μM polypeptide, the polypeptide sequence is: GTSWCYNQKRHDGP, add 1 μl acetonitrile and 6 μl 1M Tris-HCl pH 7.5, mix well;
[0069] Add 2 μl of 600 μM compound shown in formula I and mix well;
[0070] React at room temperature for 30 minutes;
[0071] Add 6.3 μl, 0.05 μl / ml 3’-mercaptopropionic acid, and terminate the reaction at room temperature for 5 minutes;
[0072] Reaction products were identified by mass spectrometry.
[0073] figure 2 It is the mass spectrogram before and after the reaction between the polypeptide and the compound shown in formula I.
[0074] The peak at 550.24924 in the mass spectrum is the polypeptide before the reaction, and the calculated molecular weight is: 1647.75. The peak a...
Embodiment 3
[0076] Embodiment 3, the fluorescent labeling of the compound shown in formula I on the protein
[0077] We selected the protein UBXD7 with a cysteine on its surface for labeling. If there is no cysteine on the surface of the protein, the surface site can also be mutated. The reaction process is as follows:
[0078] Add 5ml 20mM Tris-HCl, 150mM NaCl, pH 7.5 to the centrifuge tube, add 0.5μM UBXD7 protein, add 25μl 2mM compound, mix well;
[0079] React at room temperature for 30 minutes;
[0080] Add 250 μl, 2mM 3'-mercaptopropionic acid, and stop the reaction at room temperature for 5 minutes;
[0081] After concentration, the reaction product was identified by mass spectrometry.
[0082] image 3 It is the mass spectrogram before the reaction between UBXD7 and the compound shown in formula I, and the molecular weight of UBXD7 is: 9507.07.
[0083] Figure 4 It is the mass spectrogram after the reaction between UBXD7 and the compound shown in formula I. The molecular...
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