Authors: Czerlinski, George; Ypma, Tjalling

Source: Journal of Bionanoscience, Volume 1, Number 1, June 2007 , pp. 64-72(9)

Publisher: American Scientific Publishers

Abstract:

We investigate the feasibility of using rapid mixing and chemical relaxation by temperature jump to detect intermediates and their reaction kinetics for the telomerase catalyzed reaction on the cell nucleus. We consider only (desoxyribo-) nucleotide additions to the telomerase-telomere complex and the translocation along the extended single-stranded DNA chain. Since telomerase is not sufficiently available to conduct ordinary mixing experiments, we use 2-photon laser absorption microscopy in a volume of less than 1 μm³ (nm³-range), observing fluorescence changes due to nucleotide-binding with attached fluorophors. We distinguish between experiments on the surface of a cell nucleus and on the surface of an object glass and use both stopped flow and constant flow. Two models are considered, one incorporating a fast isomerization of the initial telomere-telomerase complex. We simulate the reactions numerically. The differences between stopped flow and constant flow are minimal. Diffusion of reactants restricts the time range available for stopped-flow experiments, so constant-flow experiments are preferred. We show that most reaction steps can be individually investigated by judicious selection of fluorophors bound to one of the three nucleotides. This mode also permits distinction between the two models. Interference by single-molecule kinetics implies that some experiments must be repeated to obtain averaged data.

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Keywords: TELOMERASE; TELOMERES; RAPID MIXING; CHEMICAL RELAXATION; TWO-PHOTON ABSORPTION

Document Type: Research article

DOI: http://dx.doi.org/10.1166/jbns.2007.007

Publication date: 2007-06-01

More about this publication?

• Bionanoscience attempts to harness various functions of biological macromolecules and integrate them with engineering for technological applications. It is based on a bottom-up approach and encompasses structural biology, biomacromolecular engineering, material science, and engineering, extending the horizon of material science. The journal aims at publication of (i) Letters (ii) Reviews (3) Concepts (4) Rapid communications (5) Research papers (6) Book reviews (7) Conference announcements in the interface between chemistry, physics, biology, material science, and technology. The use of biological macromolecules as sensors, biomaterials, information storage devices, biomolecular arrays, molecular machines is significantly increasing. The traditional disciplines of chemistry, physics, and biology are overlapping and coalescing with nanoscale science and technology. Currently research in this area is scattered in different journals and this journal seeks to bring them under a single umbrella to ensure highest quality peer-reviewed research for rapid dissemination in areas that are in the forefront of science and technology which is witnessing phenomenal and accelerated growth.
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