Mass spectrometry guided in situ proteolysis to obtain crystals for X-ray structure determination |
| |
Authors: | Tarun Gheyi Logan Rodgers Richard Romero J Michael Sauder Stephen K Burley |
| |
Institution: | 1.New York SGX Research Center for Structural Genomics, Structural Biology,Eli Lilly and Company, Lilly Biotechnology Center,San Diego,USA |
| |
Abstract: | A strategy for increasing the efficiency of protein crystallization/structure determination with mass spectrometry has been
developed. This approach combines insights from limited proteolysis/mass spectrometry and crystallization via in situ proteolysis.
The procedure seeks to identify protease-resistant polypeptide chain segments from purified proteins on the time-scale of
crystal formation, and subsequently crystallizing the target protein in the presence of the optimal protease at the right
relative concentration. We report our experience with 10 proteins of unknown structure, two of which yielded high-resolution
X-ray structures. The advantage of this approach comes from its ability to select only those structure determination candidates
that are likely to benefit from application of in situ proteolysis, using conditions most likely to result in formation of
a stable proteolytic digestion product suitable for crystallization. |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|