Naslov
Strategies for Complete Proteomic Analysis of Hydrophobic Proteins in Complex Biological Samples–Hyde-and Seek

Autori
Josic, Djuro

Izvornik
Journal of Data Mining in Genomics & Proteomics (2153-0602) 05 (2014), 01; 1-2

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, stručni

Ključne riječi
Complete Proteomic Analysis ; Hydrophobic Proteins ; Complex Biological Samples
(Proteomic Analysis ; Hydrophobic Proteins ; Complex Biological Samples)

Sažetak
The completeness of the proteomic analysis of complex biological samples frequently depends on two important factors, proper sample preparation and thorough data analysis. However, the first one is frequently neglected. The early strategies for such analyses of cells or tissues and organs started with the protein extraction out of crude homogenates or body fluids followed by proteolytic (most frequently tryptic) digestion or chemical cleavage, followed by (1 or 2D) chromatographic separation and mass spectrometric analysis of resulting peptides (LC-MS, or LC-MS/MS). Alternatively, the sample was separated by use of 1- or 2-D electrophoresis followed by excision of polypeptide bands or spots and tryptic digestion and again by LC-MS/MS analysis. However, it turned out that the performed extraction step was not complete, and a significant part of proteins in the sample were lost. Additionally, without targeted fractionation, the highly abundant as well as more hydrophilic proteins in complex samples often masked the less abundant proteins and proteins that contain hydrophobic domains and posttranslational modifications (PTMs). It frequently results in limited detection of these proteins by mass spectrometry.

Izvorni jezik
Engleski

Znanstvena područja
Biotehnologija u biomedicini (prirodno područje, biomedicina i zdravstvo, biotehničko područje)

POVEZANOST RADA