High Sensitivity Identification of Membrane Proteins by MALDI TOF-MASS Spectrometry Using Polystyrene Beads
详细信息 查看全文
- 作者：Noura Bensalem ; Sandrine Masscheleyn ; Julien Mozo ; Benoit Vallé ; e ; Franck Brouillard ; Sté ; phanie Trudel ; Daniel Ricquier ; Aleksander Edelman ; Ida Chiara Guerrera ; Bruno Miroux
- 刊名：Journal of Proteome Research
- 出版年：2007
- 出版时间：April 2007
- 年：2007
- 卷：6
- 期：4
- 页码：1595 - 1602
- 全文大小：467K
- 年卷期：v.6,no.4(April 2007)
- ISSN：1535-3907
文摘
Membrane proteins play a large variety of functions in life and represent 30% of all genomes sequenced.Due to their hydrophobic nature, they are tightly boundto their biological membrane, and detergents are alwaysrequired to extract and isolate them before identificationby mass spectrometry (MS). The latter, however remainsdifficult. Peptide mass fingerprinting methods using techniques such as MALDI-TOF MS, for example, have become an important analytical tool in the identification ofproteins. However, PMF of membrane proteins is a realchallenge for at least three reasons. First, membraneproteins are naturally present at low levels; second, mostof the detergents strongly inhibit proteases and havedeleterious effects on MALDI spectra; and third, despitethe presence of detergent, membrane proteins are unstable and often aggregate. We took the mitochondrialuncoupling protein 1 (UCP1) as a model and showed thatdifferential acetonitrile extraction of tryptic peptides combined with the use of polystirene Bio-Beads triggered highresolution of the MALDI-TOF identification of mitochondrial membrane proteins solubilized either with Triton-X100 or CHAPS detergents.