2DE-based approach for estimation of number of protein species in a cell

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 c...

Full description

Saved in:
Bibliographic Details
Published in:Electrophoresis 2014-03, Vol.35 (6), p.895-900
Main Authors: Naryzhny, Stanislav N., Lisitsa, Andrey V., Zgoda, Victor G., Ponomarenko, Elena A., Archakov, Alexander I.
Format: Article
Language:English
Subjects:
2DE
Approach
Cell
Counting
Dyes
Electrophoresis, Gel, Two-Dimensional - methods
Escherichia coli - cytology
Extrapolation
Hep G2 Cells
Histocytochemistry - methods
Human
Humans
Limit of Detection
Linear Models
Mathematical models
Number
Polypeptides
Protein species
Proteins
Proteome - analysis
Proteome - chemistry
Proteome - classification
Proteomics - methods
Spots
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:0173-0835
1522-2683
DOI:10.1002/elps.201300525