Lysosomal Fluorescent Probe

Lysosomes are organelles that break down biological macromolecules such as proteins, nucleic acids, and polysaccharides. The lysosome has a single layer membrane and various shapes. It is a bubble structure of 0.025 to 0.8 μm and contains many hydrolases. The function of lysosomes in cells is to decompose substances that enter the cells from the outside. A local cytoplasm or organelle that can digest a cell itself. When the cell ages, its lysosome ruptures, releasing a hydrolase, and digesting the entire cell to cause it to die. Lysosomes are generally an organelle in eukaryotic cells; a saclike structure coated with a single layer of membrane, the size (showing mostly spherical under electron microscopy, but having a globular sphere) of about 0.025 to 0.8 micrometers in diameter; Contains a variety of hydrolases, specifically for the decomposition of a variety of exogenous and endogenous macromolecular substances.

Figure 1. Components of a typical animal cell: 12 represents lysosome.

Characteristics

The lysosomal enzyme has three characteristics:

(1) The surface of the lysosome is highly glycosylated, which helps to protect itself from enzymatic hydrolysis. Most of the membrane proteins are glycoproteins, and the inner surface of the lysosomal membrane is negatively charged, which helps the enzyme in the lysosome to remain free. This is important for exercising normal functions and preventing cells from being digested by themselves.

(2) All hydrolases have the best activity at pH=5, but the pH value in the surrounding cytoplasm is 7.2. The lysosomal membrane contains a special transport protein, which can use the energy of ATP hydrolysis to pump H⁺ (hydrogen ion) in the cytoplasm into the lysosome to maintain its pH value=5.

(3) The enzymes in the lysosome perform their decomposition only when the hydrolyzed substance enters the lysosome. Once the lysosomal membrane is broken, the hydrolytic enzyme will escape, which will lead to autolysis of the cells.

Lysosomal Fluorescent Probe

Lysosomal fluorescent probes are a class of fluorescent dyes that selectively stain acidic compartments in living cells. These probes have several important features, 1) selective labeling of acidic organelles; 2) nanomolar (nM) The concentration can effectively mark the living cells; 3) It is provided with a multi-color probe, and the sample can be subjected to multi-standard experiments according to the situation. The lysosomal probe consists of a fluorophore group and a linked weak base. It can pass through the cell membrane freely and is generally concentrated on a spherical organelle. It is suitable for observing the internal biosynthesis of lysosomes and related pathogenesis. Lysotracker only partially protonated at neutral pH, so the principle of labeling the organelles with this probe may be related to its complete protonation and retention on the organelle membrane.

Reference:

Ohkuma S.; et al. Fluorescence probe measurement of the intralysosomal pH in living cells and the perturbation of pH by various agents. Proceedings of the National Academy of Sciences of the United States of America. 1978, 75 (7): 3327–31.

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