Highly Efficient Extraction of Cellular Nucleic Acid Associated Proteins in Vitro with Magnetic Oxidized Carbon Nanotubes
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- 作者：Yi Zhang ; Zhengyan Hu ; Hongqiang Qin ; Xiaoluan Wei ; Kai Cheng ; Fangjie Liu ; Ren鈥檃n Wu ; Hanfa Zou
- 刊名：Analytical Chemistry
- 出版年：2012
- 出版时间：December 4, 2012
- 年：2012
- 卷：84
- 期：23
- 页码：10454-10462
- 全文大小：361K
- 年卷期：v.84,no.23(December 4, 2012)
- ISSN：1520-6882
文摘
Nucleic acid associated proteins (NAaP) play the essential roles in gene regulation and protein expression. The global analysis of cellular NAaP would give a broad insight to understand the interaction between nucleic acids and the associated proteins, such as the important proteinous regulation factors on nucleic acids. Proteomic analysis presents a novel strategy to investigate a group of proteins. However, the large scale analysis of NAaP is yet impossible due to the lack of approaches to harvest target protein groups with a high efficiency. Herein, a simple and efficient method was developed to collect cellular NAaP using magnetic oxidized carbon nanotubes based on the strong interaction between carbon nanotubes and nucleic acids along with corresponding associated proteins. We found that the magnetic oxidized carbon nanotubes demonstrated a nearly 100% extraction efficiency for intracellular nucleic acids from cells in vitro. Importantly, the proteins associated on nucleic acids could be highly efficiently harvested using magnetic oxidized carbon nanotubes due to the binding of NAaP on nucleic acids. 1594 groups of nuclear NAaP and 2595 groups of cellular NAaP were extracted and identified from about 1鈥?00鈥?00 cells, and 803 groups of NAaP were analyzed with only about 10鈥?00 cells, showing a promising performance for the proteomic analysis of NAaP from minute cellular samples. This highly efficient extraction strategy for NAaP is a simple approach to identify cellular nucleic acid associated proteome, and we believed this strategy could be further applied in systems biology to understand the gene expression and regulation.