Noninvasive Analysis Of Skin Proteins In Healthy Chinese Subjects Using An Orbitrap Fusion Tribrid High-resolution Mass Spectrometer

Background In recent years,with the rapid development of disciplines such as molecular biology,biochemistry and molecular genetics,research on the physiological systems including skin has been changing at the cellular and molecular levels.Skin protein is a major component of the skin and acts as a barrier,moisturizing,signal transduction,and immune regulation in the skin barrier.Changes in skin protein content can lead to impaired skin barrier function,which is closely related to the occurrence or development of some skin diseases.The rapidly emerging mass spectrometry technology has also enabled large-scale,systematic research on proteomics.The in-depth exploration of skin proteins can provide new targets and directions for the development of new drugs and the diagnosis and treatment of skin diseases.Therefore,the function and mechanism of skin proteins have become a research hotspot.Objective The objective of this study was to perform noninvasive analysis of skin proteins in a healthy Chinese population using label-free nanoflow liquid chromatography-mass spectrometry(nLC-MS).Materials and Methods Five consecutive tape strippings were obtained from the volar forearm skin of healthy Chinese subjects.The protein on the tape was collected,extracted,digested and analyzed by a nanochromatography instrument coupled to an Orbitrap Fusion Tribrid mass spectrometer.Data-dependent acquisition allowed protein identification,which was performed by Proteome Discoverer 2.2(Thermo).Results Of the six samples in this study,we detected 114,105,122,139,129,and 151 proteins respectively,and 80 of which were common.Based on the role of these 80 proteins in the skin barrier,we divided them into 16 categories,including keratin,keratinized envelope proteins,enzymes related to cornified envelope,protease inhibitors,enzymes related to filaggrin processing,proteins related to SC cohesion,calcium binding proteins,proteins related to SC substance metabolism,protein related to antioxidant,singal transduction proteins,annexin,serum diffution markers,antimicrobial proteins,secreted protein,others,and function unkown protein.Conclusion As an important part of the skin,protein is closely related to skin physiological homeostasis and barrier function,etc.This experiment combines the non-invasive skin sample extraction method with the Orbitrap Fusion Tribrid high-resolution mass spectrometer to reduce the sampling difficulty.Improve the sensitivity of protein identification.This study provides a reference for further research on proteomics technology in human skin and disease skin.At the same time,we found in the analysis of skin proteins that there are differences in protein types between different individuals,which may be related to the inevitable errors in the process of sampling,processing and detection,and may also be different from the expression of individual skin proteins and the skin barrier status.