Study On The Interaction Between Receptor-like Kinase OsCR4 And The Extracellular Protein OsCIP1 By FRET Technology

With the deepening research of the life science,the mechanisms of various life phenomena occur,especially for the study of the interaction between the intracellular protein-protein,it becomes particularly important.In the past ten years,the proteomics develop rapidly,the interaction between proteins become the important content of proteomics.FRET(fluorescence resonance energy transfer,FRET)is a new technology developed in recent years.In 1948,it was discovered by Fosrter.FRET technology has been widely applied to the structure of biological macromolecules,especially in the immune analysis as well as quantitative analysis.In plants,transmembrane protein kinase is composed of the extracellular domain,a transmembrane region and the intracellular protein kinase domain,the protein on the one hand as a receptor for extracellular-specific ligand,at the same time is a kind of protein kinase.OsCR4 belong to the receptor kinase family.The preliminary work is based on subcellular localization analysis and in vitro activity assay,OsCR4 and extracellular signal substances may complete the process of cell differentiation.The yeast two-hybrid screen that the interaction with OsCR4,named OsCIP1.This article confirmed whether there is interaction between the two protein by FRET technology.Albumin is a carrier protein in the plasma,it can be combined with many endogenous and exogenous compounds.When they into organisms,drug through the storage and transfer of the plasma,and then they can reaches the receptor sites and play an important part in its pharmacodynamics,so it is important to study the interaction of drugs of small molecules and proteins.Scopoletin and Aesculin,are all belong to coumarin compounds,they commonly found in variety of plants,and play an important role in medical field such as anti-allergic analgesic,anti-inflammatory,anti-bacterial.This article researched rice receptor-like kinase and its extracellular protein interactions by fluorescence resonance energy transfer technology,as well as the interaction of the two kinds of coumarin medicine and bovine serum albumin,specific conclusions are as follows:1.Receptor-like kinase(OsCR4)is labeled with fluorescein isothiocyanate(FITC),and Tetramethylrhodamine-5-isothiocyanate to tag with its extracellular protein OsCIP1,fluorescence resonance energy transfer technology as the theoretical basis,we found that the two proteins in neutral buffer solutionthere is interaction with each other,when the pH of the solution is to change,they still interactions with each other.2.Bovine serum albumin(BSA)is labeled with fluorescein isothiocyanate(FITC),fluorescence resonance energy transfer technology as the theoretical basis.The experiment results are derived in the case of 1:1 molar ratio of bovine serum albumin and scopoletin,they are interaction with each other.FRET is increasing.FRET is increased with the increasing molar ratio of two substances.The interaction of the two substances is increased in different molar ratios but in the same acidity.Under the same experimental conditions,we found the results are the same,that is the bovine serum albumin is interaction with aesculin.We tried to find the combination of the saturation point of the two substances,and thus able to determine the number of binding,and So as to arrive that the mode of action between them with the BSA.Therefore the experiment to explore within a larger molar ratio and a wide pH range,but the experimental results obtained that the binding constants of the two kinds of traditional Chinese medicine and BSA are smaller,they have been combined within the scope of this experimental study.In the end,they have not reached its complexing.So the mode of action between them needs to be done to further explore.