title: Sequential extraction leading to improved proteomic analysis of the oleaginous yeast Lipomyces starkeyi.
abstract: The oleaginous yeast Lipomyces starkeyi (L. starkeyi) is an excellent intracellular lipid producer. Thus, extraction of protein from lipid-rich L. starkeyi samples following conventional methods can be difficult, leading to poor data in terms of proteomic analysis. The presence of lipophilic components in those samples may also interfere with the extraction process and the downstream analysis. In this work, we developed a sequential extraction method for preparation and analysis of L. starkeyi proteome combining to an online multidimensional nano reversed-phase liquid chromatography-tandem mass spectrometry (microRPLC-MS/MS) strategy. Protein hits of high confidence reached 227 with false positive rate less than 0.1, twice of those identified from the one-buffer extraction preparation. Moreover, the protein hits related to primary metabolism was increased, which may be important to establish the molecular mechanism of lipid accumulation. The method should be valuable for protein extraction from oleaginous species.
authors.affiliation: CAS Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Liu
authors.forename: Xin
authors.initials: X
authors.lastname: Zhao
authors.forename: Kai
authors.initials: K
authors.lastname: Cheng
authors.forename: Zongbao
authors.initials: Z
authors.lastname: Zhao
authors.forename: Mingliang
authors.initials: M
authors.lastname: Ye
mesh_terms: D005591:Chemical Fractionation / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods; D005656:Fungal Proteins / Q000032:analysis*; D055160:Lipomyces / Q000737:chemistry*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D005656:Fungal Proteins; D020543:Proteome