Protein Interaction Analysis Of Maize Endosperm AGPase-bt2and Three Crucial Glycolytic Enzymes

Corn is an important food crops and industrial raw materials. One of the hot research isabout the corn starch metabolism. Metabolic regulation of corn starch is complex biologicalprocess, completed by multiple enzymes play role in regulation. Recently, the study indicatedthat various starch metabolizing enzymes is formed multi-enzyme complex to participate in theregulation of the starch metabolism process by interaction activity in the metabolism of cornstarch process. This complex interaction of many different metabolic pathways promotesgluconeogenesis, glycolysis, starch synthesis closer to the spatial relationship of severalmetabolic pathways may indicate the existence of a common metabolic substrates, sugarglycolysis, starch metabolism pathway to promote the metabolism of starch may be through theinteraction of several proteins.Recent studies show that ADP glucose pyrophosphorylase small subunit AGPase-bt2is akey enzyme in the metabolism, and glyceraldehyde3–phosphate dehydrogenase GPN1, FBA,triosephosphate isomerization TPI enzyme are the key enzyme in the glycolytic process.Summarizes the progress of the preliminary test laboratory personnel, combined withrelevant literature reports, selecting the corn starch synthesis key enzyme ADP-glucosepyrophosphorylase small subunit AGPase-bt2, and glycolysis the key enzyme glyceraldehyde-3-phosphate dehydrogenase GPN1, fructose-bisphosphate aldolase FBA, triosephosphateisomerase TPI as experimental subjects. Our laboratory previously described the interaction ofseveral enzymes the role of yeast in the body using yeast two-hybrid, carried out to verify thework based on the use bimolecular fluorescence complementation technology (BIFC) in a largeleaf tobacco for several enzymes in vivo protein interactions were verified, and strive to restorethe true three pairs of protein interactions in plants relationships.Experimental results show that, constructed four fluorescent expression vectors, including326-CYCHA-AGPase-bt2,326-CYNEE-GPN1,326-CYNEE-FBA,326-CYNEE-TPI, and thentransformed into Agrobacterium. Using the transient expression vector conversion technologytransformed into tobacco mesophyll cells, and observed three pairs of proteins mutualinteraction situations using confocal microscopy. The results showed that three pairs of theprotein was observed were noticeable yellow signals under a microscope, and the position of therelationship interaction occurs in the chloroplasts, indicating that the interaction between threepairs of the protein in tobacco.