This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Multifunctional enzymes comprise a class of proteins that associate with each other to form enzyme complexes. Such complexes are designed so that common intermediates are channeled between active sites located distant from each other without equilibrating with the bulk phase. The advantages of such a channeling system are numerous and include such factors as preventing the loss of chemically labile intermediates as well as intermediates that are toxic to the cellular environment. The three-dimensional structures of mutifunctional enzymes play an important role in mediating the efficiency of intermediate channeling. We have solved the structure of the bifunctional enzyme 4-hydroxy-2-ketovalerate aldolase (DmpG)/acylating acetaldehyde dehydrogenase (DmpF) by MAD phasing techniques and have refined the structure to 1.7 resolution. The structure reveals a buried channel through which the toxic intermediate, acetaldehyde is channeled from the aldolase active site to the dehydrogenase active site where it is oxidized and acetylated with acetyl-CoA. Further structural studies will be carried out using X-ray diffraction data collected at SSRL. These studies are aimed at understanding the precise role of specific amino acid residues at the active sites of the two enzymes as well as residues along the buried channel. In addition we will characterize global structural changes that occur to the protein that play an important role in mediating channeling activity.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001209-27
Application #
7370354
Study Section
Special Emphasis Panel (ZRG1-BPC-E (40))
Project Start
2006-03-01
Project End
2007-02-28
Budget Start
2006-03-01
Budget End
2007-02-28
Support Year
27
Fiscal Year
2006
Total Cost
$636
Indirect Cost
Name
Stanford University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
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