Details

Autor(en) / Beteiligte

• OTSUKA, Rie
• SASAKI, Kazumi
• MISE, Miyako
• ATAKU, Hanako
• NISHIJIMA, Keiko
• YAMAZAKI, Jun
• YAMAZAKI, Syuji

Titel

Development of Anion Exchange Chromatography for Proteome Analysis and Its Application to Detection of Membrane Proteins of the Hyper-Thermophilic Crenarchaeon, Aeropyrum pernix K1

Ist Teil von

• BUNSEKI KAGAKU, 2006, Vol.55(12), pp.963-968

Ort / Verlag

Tokyo: The Japan Society for Analytical Chemistry

Erscheinungsjahr

2006

Quelle

Free E-Journal (出版社公開部分のみ）

Beschreibungen/Notizen

• For the systematic analysis of the proteomes of microorganisms whose genomic nucleotide sequences have been analyzed, a method for protein separation and purification using anion-exchange column chromatography was developed. Generally, proteome analysis has a problem that membrane proteins become insoluble during the sample preparation steps, and are difficult to detecte. We adopted the use of an anion-exchange column chromatography in the first step of protein separation, and optimized the method to allow the direct injection of whole cell lysates without any preprocessing. After removing soluble proteins, insoluble proteins retained on the column were subjected to in-column enzymatic digestion, followed by elution. The method was applied to the proteome analysis of Aeropyrum pernix K1, an aerobic hyper-thermophilic archaeon, and proteins were identified by high-performance liquid chromatography/tandem mass spectrometry (HPLC-MS/MS). Thus, a total of 10 novel membrane proteins were detected and identified. Consequently, this method seems to be effective to detect water-insoluble membrane proteins that had never been detected by other conventional methods.