title: Zn(II)-Coordinated Quantum Dot-FRET Nanosensors for the Detection of Protein Kinase Activity.
abstract: We report a simple detection of protein kinase activity using Zn(II)-mediated fluorescent resonance energy transfer (FRET) between quantum dots (QDs) and dye-tethered peptides. With neither complex chemical ligands nor surface modification of QDs, Zn(II) was the only metal ion that enabled the phosphorylated peptides to be strongly attached on the carboxyl groups of the QD surface via metal coordination, thus leading to a significant FRET efficiency. As a result, protein kinase activity in intermixed solution was efficiently detected by QD-FRET via Zn(II) coordination, especially when the peptide substrate was combined with affinity-based purification. We also found that mono- and di-phosphorylation in the peptide substrate could be discriminated by the Zn(II)-mediated QD-FRET. Our approach is expected to find applications for studying physiological function and signal transduction with respect to protein kinase activity.
authors.affiliation: Department of Life Science, Hanyang University, Seoul 133-791, Korea. plobere804@naver.com.|Research Institute for Natural Sciences, Hanyang University, Seoul 133-791, Korea. plobere804@naver.com.
authors.forename: Butaek
authors.initials: B
authors.lastname: Lim
authors.affiliation: Department of Life Science, Hanyang University, Seoul 133-791, Korea. skshzzang@nate.com.|Research Institute for Natural Sciences, Hanyang University, Seoul 133-791, Korea. skshzzang@nate.com.
authors.forename: Ji-In
authors.initials: JI
authors.lastname: Park
authors.affiliation: Department of Convergence Medicine, Asan Institute for Life Sciences, University of Ulsan College of Medicine, Asan Medical Center, Seoul 138-736, Korea. kjlee@amc.seoul.kr.
authors.forename: Kyung Jin
authors.initials: KJ
authors.lastname: Lee
authors.affiliation: Department of Life Science, Hanyang University, Seoul 133-791, Korea. jwl@hanyang.ac.kr.|Research Institute for Natural Sciences, Hanyang University, Seoul 133-791, Korea. jwl@hanyang.ac.kr.
authors.forename: Jin-Won
authors.initials: JW
authors.lastname: Lee
authors.affiliation: Department of Life Science, Hanyang University, Seoul 133-791, Korea. twgibio@hanyang.ac.kr.|Research Institute for Natural Sciences, Hanyang University, Seoul 133-791, Korea. twgibio@hanyang.ac.kr.|Research Institute for Convergence of Basic Sciences, Hanyang University, Seoul 133-791, Korea. twgibio@hanyang.ac.kr.
authors.forename: Tae-Wuk
authors.initials: TW
authors.lastname: Kim
authors.affiliation: Department of Life Science, Hanyang University, Seoul 133-791, Korea. ypilkim@hanyang.ac.kr.|Research Institute for Natural Sciences, Hanyang University, Seoul 133-791, Korea. ypilkim@hanyang.ac.kr.|Research Institute for Convergence of Basic Sciences, Hanyang University, Seoul 133-791, Korea. ypilkim@hanyang.ac.kr.
authors.forename: Young-Pil
authors.initials: YP
authors.lastname: Kim
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D015374:Biosensing Techniques / Q000379:methods*; D057075:Enzyme Assays; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D008274:Magnesium / Q000494:pharmacology; D053758:Nanoparticles / Q000737:chemistry*; D010455:Peptides / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011494:Protein Kinases / Q000378:metabolism*; D045663:Quantum Dots / Q000737:chemistry*; D013379:Substrate Specificity / Q000187:drug effects; D013997:Time Factors; D015032:Zinc / Q000737:chemistry*
chemical_list: D010455:Peptides; D000255:Adenosine Triphosphate; D011494:Protein Kinases; D008274:Magnesium; D015032:Zinc
keywords: FRET; kinase; phosphorylation; quantum dot; zinc