Production Of The TRPM8 Ion Channel In Mammalian Expression System For Structural Determination

The cold and menthol receptor,TRPM8,is a member of the transient receptor potential(TRP)cation channel family and is involved in a variety of human physiological and pathological processes,including multiple cellular functions such as pain transduction,ion homeostasis,temperature regulation,and tumorigenesis.Therefore,TRPM8 is an important target for pain and cancer.However,at present,the structure of human TRPM8 has not been resolved,which greatly hinders the understanding of the molecular mechanism of TRPM8 and the development of target drugs.To analyze the high-resolution human TRPM8 structure,this study used the mammalian expression system to screen different clones to improve protein yield,as well as different detergents to improve protein stability,and finally to obtain high-yield and high-purity human TRPM8 proteins.At the same time,we successfully assembled the TRPM8 protein in the nanodiscss,whose antigen can be used to immunize against TRPM8 related diseases such as chronic pain,cold pain and migraine.Furthermore,good biochemical and structural data indicate that we will obtain the human full-length TRPM8 structure in the short term using cryo-EM.