4OOA

CRYSTAL STRUCTURE of NAF1 (MINER1): H114C THE REDOX-ACTIVE 2FE-2S PROTEIN

Summary for 4OOA

• Entry DOI: 10.2210/pdb4ooa/pdb
• Related: 4OO7
• Descriptor: CDGSH iron-sulfur domain-containing protein 2, FE2/S2 (INORGANIC) CLUSTER (3 entities in total)
• Functional Keywords: membrane bound, thiazolidinedione, oxidative stress, metal binding protein
• Biological source: Homo sapiens (human)
• Cellular location: Endoplasmic reticulum membrane; Single-pass membrane protein: Q8N5K1
• Total number of polymer chains: 6
• Total formula weight: 46995.95

Authors

Tamir, S.,Eisenberg-Domovich, Y.,Conlan, A.R.,Stofleth, J.T.,Lipper, C.H.,Paddock, M.L.,Mittler, R.,Jennings, P.A.,Livnah, O.,Nechushtai, R. (deposition date: 2014-01-31, release date: 2014-07-02, Last modification date: 2024-02-28)

Primary citation

Tamir, S.,Eisenberg-Domovich, Y.,Conlan, A.R.,Stofleth, J.T.,Lipper, C.H.,Paddock, M.L.,Mittler, R.,Jennings, P.A.,Livnah, O.,Nechushtai, R.
A point mutation in the [2Fe-2S] cluster binding region of the NAF-1 protein (H114C) dramatically hinders the cluster donor properties.
Acta Crystallogr.,Sect.D, 70:1572-1578, 2014

PubMed Abstract: NAF-1 is an important [2Fe-2S] NEET protein associated with human health and disease. A mis-splicing mutation in NAF-1 results in Wolfram Syndrome type 2, a lethal childhood disease. Upregulation of NAF-1 is found in epithelial breast cancer cells, and suppression of NAF-1 expression by knockdown significantly suppresses tumor growth. Key to NAF-1 function is the NEET fold with its [2Fe-2S] cluster. In this work, the high-resolution structure of native NAF-1 was determined to 1.65 Å resolution (R factor = 13.5%) together with that of a mutant in which the single His ligand of its [2Fe-2S] cluster, His114, was replaced by Cys. The NAF-1 H114C mutant structure was determined to 1.58 Å resolution (R factor = 16.0%). All structural differences were localized to the cluster binding site. Compared with native NAF-1, the [2Fe-2S] clusters of the H114C mutant were found to (i) be 25-fold more stable, (ii) have a redox potential that is 300 mV more negative and (iii) have their cluster donation/transfer function abolished. Because no global structural differences were found between the mutant and the native (wild-type) NAF-1 proteins, yet significant functional differences exist between them, the NAF-1 H114C mutant is an excellent tool to decipher the underlying biological importance of the [2Fe-2S] cluster of NAF-1 in vivo.

PubMed: 24914968
DOI: 10.1107/S1399004714005458

Experimental method

X-RAY DIFFRACTION (1.58 Å)