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Scientists discover structure of protein essential for quality control, nerve function
Date:1/14/2013

-control manager for the cellular protein-making factories called ribosomes. Occasionally a ribosome receives miscoded genetic instructions and produces certain types of abnormal proteins, known as "nonstop proteins" jamming the ribosomal machinery like a wrinkled sheet of paper in an office printer. Bengtson and Joazeiro found that Ltn1 fixes jammed ribosomes by tagging nonstop proteins with ubiquitin molecules, thereby marking them for quick destruction by roving cellular garbage-disposers called proteasomes.

"The question for us then was, "How does Ltn1 do this?' " said Joazeiro.

Pushing the Boundaries of Electron Microscopy

To help find out, he began a collaboration with Carragher and Potter, who run the National Resource for Automated Molecular Microscopy (NRAMM), an advanced electron microscope facility at TSRI that is funded by the National Institutes of Health's National Center for Research Resources.

Ltn1 was deemed too large for its structure to be determined by current nuclear magnetic resonance (NMR) technology, and, as the scientists know now, too flexible to allow the highly regular crystalline packing needed by X-ray crystallographers. "It's a very floppy molecule, so it would be hard to crystallize," said Potter.

Advanced electron microscopy offered a way, however. Dmitry Lyumkis, a graduate student in the NRAMM laboratory and first author of the study, took high-resolution images of yeast Ltn1 with an electron microscope. He then used sophisticated image and data processing software to align and average individual images. The technique eliminates much of the random "noise" that obscures single images and produces a sharp 3D picture of the protein.

No one has ever used electron microscopy to distinguish so manymore than 20conformations of such a small protein. "Usually electron microscopists determine no more than two or three conformational states, and they work with protein complexes w
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Contact: Mika Ono
mikaono@scripps.edu
858-784-2052
Scripps Research Institute
Source:Eurekalert  

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