An efficient method for extraction of chloroplast protein compatible for 2-DE and MS analysis

作  者:Fan PX,Wang XC,Kuang TY,Li YX
影响因子:3.509
刊物名称:Electrophoresis
出版年份:2009
卷:30  期:1  页码:3024-3033

论文摘要:

 Comparative proteomic analysis of chloroplast by 2-DE has received significant attention
in recent years. However, the complication of membrane systems in chloroplast made it
challenging to elucidate entire chloroplast proteome by 2-DE. Here, we developed an
efficient method for extracting chloroplast proteins, and produced excellent 2-DE profiles
from both Arabidopsis thaliana and Salicornia europaea. Comparison of this method with
another two protocols for the extraction of A. thaliana chloroplast proteins showed that
our method obtained higher protein yields and produced more protein spots on both pH
3–10 and 4–7 2-DE gels. Moreover, this method recovered more proteins in the basic and
high Mr regions, thereby offering the best extraction of chloroplast proteins. Identification
of 15 specific chloroplast-targeted proteins on our gels by MALDI-TOF MS revealed
that this method was compatible with MS, and recovered more chloroplast membrane
proteins than the commonly used methods. This protocol is expected to have a wide
application in future chloroplast proteomic analysis.
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