title: Insights into the iron-ome and manganese-ome of Δmtm1 Saccharomyces cerevisiae mitochondria.
abstract: Biophysical spectroscopies and LC-ICP-MS were used to evaluate the iron-ome and manganese-ome of mitochondria from Δmtm1 yeast cells. Deleting the mitochondrial carrier gene MTM1 causes Fe to accumulate in mitochondria and Mn superoxide dismutase (SOD2) activity to decline. One explanation for this is that some accumulated Fe misincorporates into apo-Sod2p. Mössbauer spectroscopy revealed that most of the accumulated Fe was Fe(III) nanoparticles which are unlikely to misincorporate into apo-Sod2p. Under anaerobic conditions, Fe did not accumulate yet SOD2 activity remained low, suggesting that the two phenomena are independent. Mn concentrations were two-fold higher in Δmtm1 mitochondria than in WT mitochondria. Soluble extracts from such samples were subjected to size-exclusion LC and fractions were analyzed with an on-line ICP-MS. Two major Mn peaks were observed, one due to MnSod2p and the other to a Mn species with a mass of 2-3 kDa (called Mn2-3). Mn2-3 may deliver Mn into apo-Sod2p. Most Mn in WT mitochondria was associated with MnSod2p, whereas most Mn in Δmtm1 mitochondria was associated with Mn2-3. The [Mn2-3] increased in cells grown on high MnCl2 while the MnSod2p concentration remained unchanged. Corresponding Fe traces showed numerous peaks, including a complex of ~3 kDa which may be the form of Fe that misincorporates, and an Fe peak with the molecular mass of Sod2p that may correspond to FeSod2p. The intensity of this peak suggests that deleting MTM1 probably diminishes SOD2 activity by some means other than Fe misincorporation. A portion of Sod2p in Δmtm1 mitochondria might be unfolded or immature. Mtm1p may import a species required for apo-Sod2p maturation, activity or stability.
authors.affiliation: Texas A&M University, Department of Chemistry, College Station, TX 77843, USA.
authors.forename: Jinkyu
authors.initials: J
authors.lastname: Park
authors.forename: Sean P
authors.initials: SP
authors.lastname: McCormick
authors.forename: Mrinmoy
authors.initials: M
authors.lastname: Chakrabarti
authors.forename: Paul A
authors.initials: PA
authors.lastname: Lindahl
mesh_terms: D002853:Chromatography, Liquid; D005290:Ferric Compounds / Q000378:metabolism; D007501:Iron / Q000378:metabolism*; D008345:Manganese / Q000378:metabolism*; D013058:Mass Spectrometry; D008928:Mitochondria / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D015204:Spectroscopy, Mossbauer; D013482:Superoxide Dismutase / Q000378:metabolism
chemical_list: D005290:Ferric Compounds; D029701:Saccharomyces cerevisiae Proteins; D008345:Manganese; D007501:Iron; D013482:Superoxide Dismutase