High-Resolution Shadowing of Transfer RNA
AUTOR(ES)
Abermann, Reinhard J.
RESUMO
High-resolution shadowing with metals that melt at high temperatures was used to study macromolecules. Molecules of transfer RNA shadowed with tantalum-tungsten are readily visualized in an electron microscope. Mounting procedures for tRNA were perfected that reproducibly gave uniform distributions of both monomeric and dimeric tRNA particles, and allowed a statistical assessment of their gross shapes and sizes. Monomeric tRNA yielded a fairly homogeneous population of rod-shaped particles, with axial dimensions of about 40 × 85 Å. Dimers of yeast alanine tRNA held together by hydrogen bonds and dimers constructed by covalent linkage of the amino-acid acceptor (3′-) termini of monomers both gave slightly more heterogeneous populations of particles. Yet, their structures were also basically rod shaped, with their lengths ranging to about twice that of the monomer; this result indicates an end-to-end arrangement of the monomeric units within both dimers. These results suggest that the amino-acid acceptor terminus and the anticodon region are at the ends of the rod-shaped, dehydrated tRNA monomer visible by electron microscopy, consistent with the generally accepted view of tRNA structure in solution suggested by other workers using other methods. This study demonstrates that high-resolution shadowing with tantalum-tungsten provides a means to examine the three-dimensional structures of relatively small biological macromolecules.
