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Scientists develop split green for tagging protein

LOS ALAMOS, N.M., Jan. 3, 2004 - University of California scientists working at Los Alamos National Laboratory have developed a new protein tagging and detection system based on a process for "splitting" a green fluorescent protein. Unlike current protein detection methods, the method works both in living cells and in the test tube and can be used to quantify proteins down to 0.1 picomole, or one billionth of a gram of a typical protein molecule. Because the method can be used to detect protein aggregation within the living organisms, it will be useful for high-throughput studies of protein structure and protein production and for studying diseases, like Alzheimer's, that are associated with protein misfolding and aggregation.

In research published recently in the online version of the scientific journal Nature Biotechnology, Los Alamos scientists Stphanie Cabantous, Tom Terwilliger and Geoff Waldo describe a method for engineering soluble, self-associating fragments of green fluorescent proteins that can be used to tag or detect soluble and insoluble proteins in living cells or cell lysates without changing protein solubility.

According to team member Geoff Waldo, "we think this discovery will have a major impact in the field of protein biotechnology and work related to deciphering the structure and function of proteins. I like to think of it as an enabling technology, a toolbox, if you will, for protein researchers, that could help them close the gap between sequencing the DNA of the human genome and determining the structures and functions of the encoded proteins."

The new system is based on the Rapid Protein Folding Assay (RPFA) method developed several years ago by Waldo, which used green fluorescence to signal protein folding. That method worked by fusing a protein's DNA to the DNA for green fluorescent proteins (GFP). The hybrid protein created by this linking then had the characteristics of both the GFP and the protein being a
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Contact: Todd Hanson
tahanson@lanl.gov
505-665-2085
DOE/Los Alamos National Laboratory
3-Jan-2005


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