Use of Carrier Ampholyte-Free Isoelectric Focusing for Proteomic Analysis
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 phyco...
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| Published in | Chromatographia Vol. 67; no. Suppl 1; pp. 55 - 61 |
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| Main Authors | , |
| Format | Journal Article Conference Proceeding |
| Language | English |
| Published |
Wiesbaden
Wiesbaden : Vieweg Verlag
01.05.2008
Vieweg Verlag Springer |
| Subjects | |
| Online Access | Get full text |
| ISSN | 0009-5893 1612-1112 |
| DOI | 10.1365/s10337-007-0463-8 |
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| Abstract | 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. |
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| AbstractList | 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. 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. |
| Author | Procházková, Barbora Šalplachta, Jiří |
| Author_xml | – sequence: 1 givenname: Barbora surname: PROCHAZKOVA fullname: PROCHAZKOVA, Barbora organization: Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University of Agriculture and Forestry Brno, Zemědělská 1, 613 00 Brno, Czech Republic – sequence: 2 givenname: Jirri surname: SALPLACHTA fullname: SALPLACHTA, Jirri organization: Institute of Analytical Chemistry, Academy of Sciences of the Czech Republic, Veveri 97, 602 00 Brno, Czech Republic |
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| Keywords | 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 |
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| SubjectTerms | 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 |
| Title | Use of Carrier Ampholyte-Free Isoelectric Focusing for Proteomic Analysis |
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