This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.The nucleus, mitochondria and vacuolar system are cellular compartments containing subsets of proteins that carry out specific functions. We propose to determine the form of metals and metalloids localized to compartments using two systems. First, we propose to determine the form of accumulated iron in isolated mitochondria. Mitochondrial iron overload is a common feature of several neurological and metabolic diseases. Currently, it is assumed that all excess mitochondrial iron is the same but there is no structural evidence to support this view. The form of iron that accumulates may depend on the pathway that is disrupted. To address this issue we will utilize the well-characterized yeast iron/sulfur cluster pathway. Using XAS, we propose to characterize the mitochondrial iron that accumulates when specific proteins, linked to iron/sulfur cluster assembly, are mutated. We will collect iron K-edges for fingerprinting and examine both the near edge and the EXAFS when appropriate. Since some of these yeast genes have human homologues that cause specific diseases, a definitive characterization of the species present could lead to the development of 'custom iron chelators'. Secondly, we propose to determine copper and selenium species in whole animals and organs that are known to sequester these compounds. X-ray absorption spectroscopy imaging is a relatively new technique that has been used to interrogate the spatial distribution of different chemical species in situ. We propose to extend this technique to two animal models, the compartmentalization of copper in the cuprophilic cells of the fruit fly and selenium bioaccumulation in the water boatman. Both Se and Cu XAS spectra, have a difference of about 7eV between the near edge maximal peak positions of selenite vs selenate and cupric vs cuprous compounds. This shift provides ample contrast for chemically selective XAS imaging. Our long-term goal is to apply XAS imaging to mammalian systems.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001209-28
Application #
7597951
Study Section
Special Emphasis Panel (ZRG1-BPC-E (40))
Project Start
2007-03-01
Project End
2008-02-29
Budget Start
2007-03-01
Budget End
2008-02-29
Support Year
28
Fiscal Year
2007
Total Cost
$2,181
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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