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MPP OUTSIDE REQUEST FORM 12/1/10 Requestor(s ): Elizabeth Craig Institution: UW Biochemistry Request Title: yeast mitochondrial Nfu1. Sequence alignment and domain structure of Nfu1p of S. cerevisiae. 26. Signal sequence. 1. 122. FeS binding site.
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MPP OUTSIDE REQUEST FORM 12/1/10 Requestor(s): Elizabeth Craig Institution: UW Biochemistry Request Title: yeast mitochondrial Nfu1
Sequence alignment and domain structure of Nfu1p of S. cerevisiae 26 Signal sequence 1 122 FeS binding site 243 256 : N-terminal domain, 22-123 aa. (Human NFU N-termnal domain: structured domain) : Nfu-like domain, 148-230 aa. (Human NFU C-teminal domain: molten globule domain, flexible)
10/22/11 Update – Our partner the NorthEast Structural Genomics consortium (NESG) has purified many samples of S. cereviseae Nfu1 truncations and obtained good NMR HSQC spectra on some of them, making a structure or structures in the near future quite likely. The NESG code for yeast Nfu1 is YR313; domains are named YR313A, B, etc, and series of truncations around those domains were constructed. The most successful are: YR313A, residues 17-124: ~137 mg purified, labeled with 15N/13C-5%, good HSQC obtained YR313A, residues 24-130, ~89 mg purified, labeled with 15N/13C-5%, good HSQC obtained Full-length protein has also expressed well and solubly