Detection of CREB phosphorylation via Zr (IV) ion mediated signal amplification.
Phosphorylation of protein plays a vital regulatory role in a variety of biological processes. We herein report a novel method to assay the level of phosphorylated cAMP-response element binding protein (CREB) via Zr(4+) mediated signal amplification using gold nanoparticle/DNA/methylene blue (GNP/DNA/MB) nanocomposites. In this method the probe DNA immobilized at a gold electrode surface can specifically and efficiently recognize the phosphorylated target protein CREB. Then Zr(4+) links the phosphorylated CREB with GNP/DNA/MB nanocomposites by coordinating the phosphate groups on both CREB and the nanocomposites. Since the nanocomposites can provide high sensitivity (limit of detection: 0.25 nM) for the detection, efficient and highly sensitive bioanalysis of the expression level of phosphorylated protein CREB in human placenta tissues has also been conducted in this work. Our method is reported which shows acceptable stability, reproducibility for assaying of the protein phosphorylation states in real biosamples under physiological and pathological conditions with great potential for clinical applications in future.
Department of Obstetrics, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, PR China; Department of Biochemistry and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PR China.
Tingting
T
Yin
Department of Biochemistry and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PR China.
Hao
H
Li
Department of Obstetrics, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, PR China; Department of Biochemistry and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PR China.
Nana
N
Yang
Department of Biochemistry and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PR China.
Tao
T
Gao
Department of Obstetrics, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, PR China. Electronic address: lizhou_sun121@hotmail.com.
Lizhou
L
Sun
Department of Biochemistry and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PR China; Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, PR China. Electronic address: genxili@nju.edu.cn.
Genxi
G
Li
D015374:Biosensing Techniques / Q000379:methods*; D002412:Cations / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D017362:Cyclic AMP Response Element-Binding Protein / Q000032:analysis / Q000378:metabolism*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D055664:Electrochemical Techniques / Q000379:methods*; D005260:Female; D006046:Gold / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D057230:Limit of Detection; D053758:Nanoparticles / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D015203:Reproducibility of Results; D015040:Zirconium / Q000378:metabolism*
D002412:Cations; D017362:Cyclic AMP Response Element-Binding Protein; D006046:Gold; D004247:DNA; D015040:Zirconium; D000242:Cyclic AMP
DNA/protein interaction; Gold nanoparticle; Protein phosphorylation; Square wave voltammetry; cAMP-response element binding protein