Use of Carrier Ampholyte-Free Isoelectric Focusing for Proteomic Analysis

• Published in: Chromatographia Vol. 67; no. Suppl 1; pp. 55 - 61
• Main Authors: PROCHAZKOVA, Barbora, SALPLACHTA, Jirri
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Wiesbaden Wiesbaden : Vieweg Verlag 01.05.2008; Vieweg Verlag; Springer
• Subjects: Analytical Chemistry; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chemistry; Chemistry and Materials Science; Chromatography; Fundamental and applied biological sciences. Psychology; isoelectric focusing; Laboratory Medicine; mass spectrometry; microalgae; nitrogen; Non metallic chromoproteins, photoproteins; Original; Pharmacy; phycobiliprotein; Porphyridium cruentum; Proteins; proteolysis; Proteomics; spectrometers; trypsin; ultrasonics; Phycobiliproteins; Protein micropreparation; Matrix assited laser desorption ionization-time-of-flight-TOF-mass spectrometry; Carrier ampholyte-free isoelectric focusing; Proteomics; Phycocyanin; Tandem mass spectrometry; Chemical analysis; Peptides; Matrix assisted laser desorption ionization; Matrix assited laser desorption ionization-time-of-flight-TOF-mass spectrometry;Carrier ampholyte-free isoelectric focusing;Proteomics;Protein micropreparation;Phycobiliproteins; Protein; Chemical enrichment; Separation method; Analysis method; Microbial origin; Plankton; Peptide fragment; Time of flight method; Cyanobacteria; Phycoerythrin; Bacteria; Phytoplankton; Biliprotein; Isoelectric focusing
• ISSN: 0009-5893; 1612-1112
• DOI: 10.1365/s10337-007-0463-8

Carrier ampholyte-free isoelectric focusing was applied for pre-concentration, purification and micropreparation of phycobiliproteins (C-phycocyanin, allophycocyanin, B-phycoerythrin) extracted from cyanobacteria Anabeana doliolum and from red microalga Porphyridium cruentum. The extraction of phycobiliproteins was carried out in deionized water. The sonication in the ultrasonic bath and liquid nitrogen freeze grind was used for extraction of proteins of interest. Pre-concentrated and pre-separated proteins were collected and analyzed via MALDI-TOF-TOF mass spectrometer after their proteolytic digestion via trypsin. Based on tandem mass spectrometric analysis, the C-phycocyanin, allophycocyanin and B-phycoerythrin were identified unambiguously.