2 research outputs found
Virtual-Experimental 2DE Approach in Chromosome-Centric Human Proteome Project
To
obtain more information about human proteome, especially about
proteoforms (protein species) coded by 18th chromosome, we separated
proteins from human cancer cell line (HepG2) by two-dimensional gel
electrophoresis (2DE). Initially, proteins in major spots were identified
by MALDI-MS peptide mass fingerprinting. According to parameters (p<i>I</i>/<i>M</i><sub>w</sub>) of identified proteins
the gel was calibrated. Using this calibrated gel, a virtual 2D map
of proteoforms coded by Chromosome 18 was constructed. Next, the produced
gel was divided into 96 sections with determined coordinates. Each
section was cut, shredded, and treated by trypsin according to mass-spectrometry
protocol. After protein identification by shotgun mass spectrometry
using ESI LC–MS/MS, a list of 20 462 proteoforms (product
of 3774 genes) was generated. Among them, 165 proteoforms are representing
39 genes of 18th chromosome. The 3D graphs showing the distribution
of different proteoforms from the same gene in 2D map were generated.
This is a first step in creation of 2DE-based knowledge database of
proteins coded by 18th chromosome
Virtual-Experimental 2DE Approach in Chromosome-Centric Human Proteome Project
To
obtain more information about human proteome, especially about
proteoforms (protein species) coded by 18th chromosome, we separated
proteins from human cancer cell line (HepG2) by two-dimensional gel
electrophoresis (2DE). Initially, proteins in major spots were identified
by MALDI-MS peptide mass fingerprinting. According to parameters (p<i>I</i>/<i>M</i><sub>w</sub>) of identified proteins
the gel was calibrated. Using this calibrated gel, a virtual 2D map
of proteoforms coded by Chromosome 18 was constructed. Next, the produced
gel was divided into 96 sections with determined coordinates. Each
section was cut, shredded, and treated by trypsin according to mass-spectrometry
protocol. After protein identification by shotgun mass spectrometry
using ESI LC–MS/MS, a list of 20 462 proteoforms (product
of 3774 genes) was generated. Among them, 165 proteoforms are representing
39 genes of 18th chromosome. The 3D graphs showing the distribution
of different proteoforms from the same gene in 2D map were generated.
This is a first step in creation of 2DE-based knowledge database of
proteins coded by 18th chromosome