title: [Experimental estimation of proteome size for cells and human plasma].
abstract: Huge range of concentrations of different protein and insufficient sensitivity of methods for detection of proteins at a single molecule level does not yet allow obtaining the whole image of human proteome. In our investigations, we tried to evaluate the size of different proteomes (cells and plasma). The approach used is based on detection of protein spots in 2-DE after staining by protein dyes with different sensitivities. The function representing the dependence of the number of protein spots on sensitivity of protein dyes was generated. Next, by extrapolation of this function curve to theoretical point of the maximum sensitivity (detection of a single smallest polypeptide) it was calculated that a single human cell (HepG2) may contain minimum 70,000 proteoforms, and plasma--1.5 mln. Utilization of this approach to other, smaller proteomes showed the competency of this extrapolation. For instance, the size of mycoplas ma (Acholeplasma laidlawii) was estimated in 1100 proteoforms, yeast (Saccharomyces cerevisiae)--40,000, E. coli--6200, P. furiosus--3400. In hepatocytes, the amount of proteoforms was the same as in HepG2--70,000. Significance of obtained data is in possibilities to estimating the proteome organization and planning next steps in its study.
authors.affiliation: Institute of Biomedical Chemistry, Moscow, Russia; Konstantinov Petersburg Nuclear Physics Institute, Gatchina, Leningrad District, Russia.
authors.forename: S N
authors.initials: SN
authors.lastname: Naryzhny
authors.affiliation: Institute of Biomedical Chemistry, Moscow, Russia.
authors.forename: V G
authors.initials: VG
authors.lastname: Zgoda
authors.affiliation: Institute of Biomedical Chemistry, Moscow, Russia.
authors.forename: M A
authors.initials: MA
authors.lastname: Maynskova
authors.affiliation: Konstantinov Petersburg Nuclear Physics Institute, Gatchina, Leningrad District, Russia.
authors.forename: N L
authors.initials: NL
authors.lastname: Ronzhina
authors.affiliation: Konstantinov Petersburg Nuclear Physics Institute, Gatchina, Leningrad District, Russia.
authors.forename: N V
authors.initials: NV
authors.lastname: Belyakova
authors.affiliation: Konstantinov Petersburg Nuclear Physics Institute, Gatchina, Leningrad District, Russia.
authors.forename: O K
authors.initials: OK
authors.lastname: Legina
authors.affiliation: Institute of Biomedical Chemistry, Moscow, Russia.
authors.forename: A I
authors.initials: AI
authors.lastname: Archakov
mesh_terms: D000128:Acholeplasma laidlawii / Q000166:cytology / Q000378:metabolism; D001798:Blood Proteins / Q000032:analysis*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004926:Escherichia coli / Q000166:cytology; D029968:Escherichia coli Proteins / Q000032:analysis; D005456:Fluorescent Dyes; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D057230:Limit of Detection; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis
chemical_list: D001798:Blood Proteins; D029968:Escherichia coli Proteins; D005456:Fluorescent Dyes; D020543:Proteome; D029701:Saccharomyces cerevisiae Proteins
keywords: 2DE; proteoforms; proteome