Funktionsbasierte Isolierung von Subproteomen mit trifunktionellen Fangverbindungen

Isolation, identification and quantitation of functional subproteomes by use of trifunctional capture compounds™, followed by mass spectrometric analysis, is a relatively new, promising technology for proteomic research. Particularly for analysis of effects and side effects of drugs or plant protection agents, as well as for the molecular biological basic research, the capability of this technology could be immense. When in 2005 the first photoaktivable capture compound™ for methyltransferases was avaiable at Aachen University, it was still unclear how it should be implemented to obtain samples suitable to further mass spectrometric analysis. An important challenge during this work was therefore, working out suitable basic conditions. This included the checkout of a suitable pretreatment of test material, viable buffer terms and efficient photo activation, as well as the maximization of the yield in specific proteins and a minimization of non-specific proteins. As a second aim, the new technology should be tested with the model organism E. coli. This, at the one hand, was a good demonstration of her applicability and, at the other hand, posed a challenge in that many of the target proteins thitherto have never been isolated. Beyond that, these rare expressed target proteins do not show a high affinity towards their cofactor and the used capture compound™. E. colis “methylome” therefore was a fascinating test object to check out the new methods capabilities and enabled a fair comparison to other up to date techniques.

Grantor dc:publisher

Publikationsserver der RWTH Aachen University

Year dc:date

2011

Author dc:creator

• Poot, Peter

Contributors dc:contributor

• Weinhold, Elmar

• info:eu-repo/classification/ddc/570
• Transferasen
• Methyltransferasen
• Adenosylmethionin
• Proteomanalyse
• LC-MS
• Proteine
• DNS-Methyltransferase
• Escherichia coli
• Biowissenschaften, Biologie
• Methyltransferase
• Fangverbindung
• SAM
• SAH
• proteomics
• capture compound
• AdoMet
• AdoHcy

Statement dc:rights

• info:eu-repo/semantics/openAccess

Language dc:language

ger

Identifier

https://publications.rwth-aachen.de/search?p=id:%22RWTH-CONV-114577%22

OAI record oai:identifier

https://publications.rwth-aachen.de/oai2d?verb=GetRecord&metadataPrefix=oai_dc&identifier=oai%3Apublications.rwth-aachen.de%3A52346