Methods, systems, and compositions for studying solvent accessibility and three-dimensional structure biological molecules
A technology of biomolecules and solvents, applied in the field of modifying multiple biomolecules located in multiple samples, and modifying biomolecules located in samples, which can solve problems such as expensive equipment and expensive
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Embodiment 1
[0141] Example 1. Labeling Cytochrome C
[0142] like figure 1 The system shown is used to generate free radicals in protein solutions. The plasma is prepared as follows: A low voltage AC power supply generates a variable frequency signal in the audio frequency range. This signal was fed into a Trek amplifier (commercially available from Trek Corporation, Lockport, NY) to generate a high voltage signal of the same frequency up to 30 kHz. The output of the Trek amplifier was then fed into a nickel needle that was placed above the protein solution in the glass tube. The glass tube is 0.75 inches in height and 0.5 inches in diameter. The tube was sealed to a glass foil (0.0625 inches thick), which served as a dielectric barrier. Place the copper electrode on the opposite side of the glass dielectric and connect to the other side (ground) of the signal generator. Plasma is formed in microsecond pulses whenever the voltage between the needle and copper electrode exceeds a cert...
Embodiment 2
[0145] Example 2. Labeling bovine serum albumin
[0146] like figure 1 The system shown was used to generate free radicals in protein solutions using the same operating parameters as described in Example 1.
[0147] Purified bovine serum albumin (BSA) was tested in mildly buffered saline at a concentration of 10 μΜ under the following conditions: no plasma exposure; 30 seconds plasma exposure; and 1 minute plasma exposure. Four different peptides within BSA were modified in a dose-dependent manner by this method.
[0148] refer to Image 6 , shows the percent modification of four different peptides under three different conditions. In conjunction with Example 1, this example illustrates the effectiveness of the system and method in labeling proteins of different concentrations, proteins of different sizes and proteins with different physiochemical properties.
Embodiment 3
[0149] Example 3. Size-dependent and Exposure-Dose-Dependent Forms of DNA Breakdown
[0150] Purified phage lambda genomic DNA samples in water were used as samples, using a method similar to figure 1 The experimental setup is shown and the experimental parameters described in Example 1. The lambda phage genome contains 48,500 base pairs and is linear. Samples were exposed to no plasma, plasma for 5 seconds, 10 seconds, 15 seconds, 30 seconds, 45 seconds and 60 seconds. Figure 7 An electrophoresis gel run with DNA samples after plasma exposure is shown. The samples with no exposure, 5 sec exposure and 10 sec exposure were essentially unmodified. Samples exposed for 15 seconds, 30 seconds, 45 seconds and 60 seconds produced tailing in the gel. Tailing represents a number of different DNA breakdown products resulting from the non-specific cleavage of DNA by hydroxyl radicals, thus resulting in fragments of various sizes. The tailing migration that occurred with increasing ...
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