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2DE‐based approach for estimation of number of protein species in a cell
Author(s) -
Naryzhny Stanislav N.,
Lisitsa Andrey V.,
Zgoda Victor G.,
Ponomarenko Elena A.,
Archakov Alexander I.
Publication year - 2014
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201300525
Subject(s) - proteome , extrapolation , function (biology) , escherichia coli , protein detection , biology , spots , sensitivity (control systems) , human proteome project , proteomics , chemistry , computational biology , biochemistry , microbiology and biotechnology , nanotechnology , mathematics , mathematical analysis , materials science , botany , electronic engineering , gene , engineering
Insufficient sensitivity of methods for detection of proteins at a single molecule level does not yet allow obtaining the whole image of human proteome. But to go further, we need at least to know the proteome size, or how many different protein species compose this proteome. This is the task that could be at least partially realized by the method described in this article. The approach used in our study is based on detection of protein spots in 2DE after staining by protein dyes with various sensitivities. As the different protein spots contain different protein species, counting the spots opens a way for estimation of number of protein species. The function representing the dependence of the number of protein spots on sensitivity or LOD of protein dyes was generated. And extrapolation of this function curve to theoretical point of the maximum sensitivity (detection of a single smallest polypeptide) allowed to counting the number of different molecules (polypeptide species) at the concentration level of a single polypeptide per proteome. Using this approach, it was estimated that the minimal numbers of protein species for model objects, Escherichia coli and Pirococcus furiosus , are 6200 and 3400, respectively. We expect a single human cell (HepG2) to contain minimum 70 000 protein species.