How is the level of phosphorylation determined?

The level of phosphorylation of organic molecules or substances is determined by two methods: western blotting and weight spectrometry. Phosphorylation most often refers to the connection of phosphate to protein. When phosphates are connected to anti -ns, natural processes and function of protein enzymes in the human body are disrupted. Determination of phosphorylation is essential for the treatment of a wide range of disease, from cancer to diabetes. Antibodies, such as monoclonal antibodies against anti-phosphosspho-tyrosine, are usually used because they are easily bound to the remains of tyrosine protein that are affected by phosphorylation. In western transmission, proteins are first separated by the size by means of an electric charge known as gel electrophoresis.es on proteins. Once proteins are transferred, antibodies will stick to any proteins with connected phosphate. The level of phosphorylation can then be determined by the X -ray of the membrane. Any affected proteins will release a flash of light, which will make it easier for the researcher to measure levelsphosphorylation.

Weight spectrometry is another way to measure the level of phosphorylation of the substance. This process converts protein molecules into ions, making it easier to monitor and measure. The sample is ionized by a source of high -performance ions that flood the electrons sample. This event changes the sample into cations, positively charged ions.

The material analyzer of the mass spectrometer separates the ions of their weight and the charge intensity. This allows the researcher to discover the weight ratio to the charge, which is decisive in discovering the phosphorylation level. Finally, the detector measures the ions in order to show the data in the form of a graph. The graph shows the weight ratio to the ion hub on the X axis and the relative intensity of the ions on the Y axis.

Phosphorylation levels

can also be measured during substrate phosphorylation. The phosphorylation of the substrate is a process that creates adenosine triphosphate (ATP) during glycolysisOU for energy transfer between cells. This process occurs both in animal cells and in plant cells.

In plant cells, ATP molecules are created by energy from the sun. This energy causes phosphorylation in chloroplasts of plants, at the site of photosynthesis of plants. The phosphorylation level correlates with the amount of phosphate needed to create other molecules, etc.