title: Isolation technique and proteomic analysis of the erythrocyte ghosts of red-eared turtle (Trachemys scripta).
abstract: To proceed proteomic analysis of erythrocyte of the red-eared turtle Trachemys scripta, a method for obtaining turtle erythrocyte ghosts (TEG) was first developed. After hypotonic lysis using a special buffer, forcing the erythrocyte through the syringe with an "N"-shaped needle, applying low speed homogenizing and differential centrifugation, highly purified TEG fractions were obtained. The isolated TEG proteins were treated with in-gel digestion separated by SDS-PAGE or in-solution digestion followed by HPLC predissociation, and then identified by nano-ESI-LC MS/MS techniques. A total of 169 TEG proteins was identified, validated, and categorized. Among these proteins, tubulins, and cell-surface-located F-type ATP synthase revealed important information into the super tolerance of Trachemys scripta in anoxia and low temperature exposure. Altogether, this study not only provided a method to isolate high quality TEG and a dataset of comprehensive characterization of TEG proteins, but also provides a tool for proteomic research of all nucleated red blood cells, and thus opened a new research field for exploring the mechanisms of super tolerance of turtles in harsh environment.
authors.affiliation: College of Life Sciences, Hunan Normal University, Changsha, Hunan, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Gao
authors.forename: Jianglin
authors.initials: J
authors.lastname: Li
authors.forename: Can
authors.initials: C
authors.lastname: Feng
authors.forename: Zhaotun
authors.initials: Z
authors.lastname: Hu
authors.forename: Wenfeng
authors.initials: W
authors.lastname: Liu
authors.forename: Songping
authors.initials: S
authors.lastname: Liang
authors.forename: Dazhong
authors.initials: D
authors.lastname: Yin
mesh_terms: D000818:Animals; D001798:Blood Proteins / Q000032:analysis / Q000737:chemistry; D002469:Cell Separation / Q000379:methods*; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000737:chemistry*; D020543:Proteome / Q000032:analysis / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry; D014426:Turtles / Q000097:blood*
chemical_list: D001798:Blood Proteins; D020543:Proteome