PGK is the key enzyme of glycolysis. It's also a key enzyme for organisms to survive. It widely exists in
animals, plants and microorganisms. It has many biological functions, such as affecting DNA replication
and repair, stimulating RNA synthesis of virus, and is widely used in drug target design.
PGK catalyzes the production of 1,3-diphosphoglyceride and ADP from 3-phosphoglycerate and ATP. 1,3-
diphosphoglyceride produces 3-phosphoglyceraldehyde, NAD and phosphoric acid under the action of 3-
phosphoglyceraldehyde dehydrogenase and NADH. The absorbance decreased at 340nm. It reflects the
activity of 3-phosphoglycerate kinase.
Required but Not Provided:
Ultraviolet spectrophotometer/microplate reader, desk centrifuge, water-bath, transferpettor, micro quartz
cuvette/96 well UV plate, mortar/homogenizer, ice and distilled water.
Protocol
I. Preparation:
1. Tissue: according to the tissue weight (g): the volume of the Extract solution (mL) is 1:5-10. It is
suggested that add 1 mL of Extract solution to 0.1 g of tissue. Homogenate on ice. Centrifuge at 10000 g
4℃ for 10 minutes. Take the supernatant on ice for test.
2. Cells: according to the number of the cells (104): the volume of the Extract solution (mL) is
500~1000:1. It is suggested that add 1 mL of Extract solution to 5 million of cells. Breaking cells by