Regulation Mechanism Of HIV Membrane Protein Inhibitors C34 By Red Light

AIM: Acquired immunodeficiency syndrome(AIDS)is a serious infectious disease caused by human immunodeficiency virus(HIV).The HIV life cycle is generally divided into several different stages: virus adsorption,binding and fusion,virus shelling,transcription and translation,integration,assembly,and budding of mature.It is very important to understand the details of the course of HIV infection,but some of them are so fast that it is difficult to know what is happening at a time.The research direction of this subject is the fusion stage of virus invading target cells.METHODS: Gp41 is the envelope protein complex of HIV-I,which controls the early critical process of viral invasion into the target cells.C34 is a synthetic peptide in the core CHR region,which can inhibit the process of HIV-I fusion by inhibiting the binding of C34 peptide of HIV-I gp41 and N36 peptide of NHR region.Many anti HIV drugs on the market now apply this principle.Here,we developed an open and closed system,which can use light to control the HIV into the target cell membrane process.The structure and function of azobenzene photoisomerization provides an important molecular tool for us.Aminoazobenzene response with the strong light,the characteristic of which shows great potential for development in many fields.There are usually azobenzene CIS and trans isomers of two.Trans azobenzene into CIS configuration,need to UV light in a specific wavelength under visible light or heat;in effect,can be reduced to CIS trans configuration.Two configurations of UV-visible absorption spectrum of the azobenzene molecules are significantly different.At the same time,there are obvious differences in the physical and chemical properties of the three-dimensional structure,dipole moment and so on.At present,the different characteristics of azobenzene CIS trans isomers,CIS trans isomerization and induced light response phenomenon,has aroused widespread concern in the community.Due to UV to cells and tissues has strong killing effect,so the key to in vivo application of azobenzene is the need to provide a suitable wavelength for the isomerization reaction of the irradiation.Our p-azophenyl is modified,the regulation of wavelengths in red region(630-660 nm).The key features of the switch system is to modify the C34 peptide,through the design of C34 to cysteine mutants crosslinked azobenzene and design optical redshift of the position.Here we designed 6 mutant polypeptide and two optical switch.The synthesis of peptides was carried out by solid phase synthesis method.The purification of the peptides was performed by preparative high performance liquid chromatography.The structure of the peptide was identified by MALDI-TOF method,and the purity was determined by analytical high performance liquid chromatography(HPLC).Optical switch by chemical synthesis and purification were purified by silica gel column chromatography,using MALDI-TOF and 1H NMR spectrum analysis,the purity was determined by HPLC;using UV spectrophotometer scanning optical switch ultraviolet visible spectrum.The activity of the switch system was determined by the method of cell fusion,and the structure of the system was determined by the circular two chromatographic scanning method.RESULTS: 1.The key to the success of the experiment is the synthesis and purification of peptides.The experiment was completed successfully,and it was proved to be accurate and pure by mass spectrometry and HPLC analysis.2.the synthesis of the switch is more complex,after repeated experiments and optimization,the final product yield is high,high purity.The results of mass spectrometry and HPLC analysis showed that the structure of the product was accurate and the purity was high.3.the assembly of the switch system is relatively simple,the product obtained by mass spectrometry and HPLC analysis of the product structure is accurate,high purity,in line with the requirements of the experiment.4.circular chromatography(CD)scan can be seen that the switch system has two more obvious alpha spiral slot,which is also consistent with the C peptide and N peptide formed after the binding of the helix of the polymer structure of the six.Through the comparison of C34 and its mutant CD spectrum,we can see that the mutation of amino acid has some interference to the formation of helix.However,the peak pattern was consistent with the C34 control group.5.The results showed that the activity of the mutant was decreased,and the activity of C34-M3 was lost.The same results as CD can be seen by red and blue.C34-M4 red light is less than twice as much as that of blue light,and the red light of C34-a is less than twice that of blue light.CONCLUSION: the isomerization wavelength of the switching system moves towards the long wavelength(red linght),which is also consistent with the original design of the switch.Red light can penetrate biological cells and tissues and organs,no harm to tissues and organs.This makes the switch system more suitable for the study of living organisms.We are the light control function test switch system: some exhibited inhibitory activity of HIV in an after blue light irradiation of them,in the red light irradiation will lose the inhibition function.