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Enzyme detectives uncover new reactions, products
Date:9/8/2008

shift in double-bond position, the enzyme modification had yielded three completely new products -- two variations of a hydroxylated product called an allylic alcohol and a fatty acid containing two double bonds. "This was a profound shift in enzyme function," noted Shanklin, who has been working with modified enzymes for 15 years. "Usually you make changes very gradually, getting a few percent of a new product mixed with the original product. This was more like throwing a switch, making the change in function close to complete."

The discovery is also notable because the starting enzyme, like other soluble (membrane-independent) desaturases, can ordinarily perform only its one specified reaction -- desaturation. This is unlike desaturase enzymes that reside within the cell membrane, which appear to be more versatile, performing a range of reactions. The soluble and membrane enzymes, however, do share one key feature: both perform reactions that require the production of a highly reactive form of oxygen.

"Since both classes of enzymes produce activated oxygen, in theory the soluble enzymes, like their membrane counterparts, should be able to perform a variety of reactions as well," Shanklin said. "Our work demonstrates that this is indeed the case. Making small changes to the enzyme's amino acid sequence has unlocked the soluble desaturase's potential to facilitate a wider range of chemistry than has been seen before," Shanklin said.

The challenge is to figure out how these structural changes to the enzyme lead to the observed changes in reaction chemistry. Computer-generated models combining the known structure of the starting enzyme in conjunction with its new substrates are helping the scientists understand how the enzyme works. The next step is to obtain real 3-D crystal structures of enzyme-substrate complexes, using the National Synchrotron Light Source at Brookhaven Lab, to see how they match up with the predictions.

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Contact: Karen McNulty Walsh
kmcnulty@bnl.gov
631-344-8350
DOE/Brookhaven National Laboratory
Source:Eurekalert

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