Title :
RNA labeling and immobilization for nano-displacement measurement: probing three-dimensional RNA structure
Author :
Akabayov, Barak ; Henn, Arnon ; Elbaum, Michael ; Sagi, Irit
Author_Institution :
Dept. of Struct. Biol., Weizmann Inst. of Sci., Rehovot, Israel
fDate :
6/1/2003 12:00:00 AM
Abstract :
RNA molecules play essential roles in many biological processes, including the storage and transfer of information in the cell. These events are mediated via RNA-protein interactions or by catalytic RNA molecules. It is now recognized that unique RNA folds are associated with biological functions. Therefore, to study the intrinsic structural changes and dynamics which regulate the various functions of RNA, it is necessary to probe its three-dimensional structure in solution. In this respect, using single-molecule methodologies may allow study of native RNA molecules independent of their size and in real time. However, this may require the immobilization of RNA on a surface. Here, we report a novel approach to immobilize RNA on a glass. The procedures involve both chemical and enzymatic modifications of long RNA molecules. In addition, we demonstrate the application of an optical tweezers apparatus to measure the length and, hence, the dynamics of immobilized intact ribosomal RNA molecules as a function of different solution conditions.
Keywords :
biological techniques; displacement measurement; macromolecules; molecular biophysics; molecular configurations; RNA immobilization; RNA labeling; bead coating; biophysical technique; chemical modifications; enzymatic modifications; long RNA molecules; nanodisplacement measurement; optical tweezers; single-molecule methodologies; solution conditions; three-dimensional RNA structure probing; Biological processes; Biomedical optical imaging; Cells (biology); Chemicals; Glass; Labeling; Length measurement; Nanobioscience; Probes; RNA; Adsorption; Feasibility Studies; Glass; Magnesium Chloride; Micromanipulation; Motion; Nanotechnology; Nucleic Acid Conformation; Optics; RNA; RNA, Ribosomal, 23S; Staining and Labeling;
Journal_Title :
NanoBioscience, IEEE Transactions on
DOI :
10.1109/TNB.2003.813937