While the iron-containing photosynthetic proteins ferredoxin (Fd) and cytochrome f (Cyt f) were already decreased 75% in iron-deficient (1-μM Fe) relative to iron-replete photoheterotrophic

cells, phototrophic cells retained their iron-containing proteins until severely iron-limited conditions (0.1-μM Fe). To establish that the decrease in abundance of iron-containing proteins is a specific response to iron deficiency rather than to growth inhibition, we monitored the abundance of Fe-independent proteins LhcSR and ferroxidase (Fox1) whose expression increases in iron-deficient cells (La Fontaine et al. 2002; Naumann et al. 2007). Indeed, the expression of Fox1, a marker of Fe-deficiency, was reciprocal to the abundance of Fe-containing

photosynthetic proteins (Fig. 7). #Selleckchem GDC0068 randurls[1|1|,|CHEM1|]# The abundance of LhcSR, which is necessary for NPQ (Peers et al. 2009), increased with respect to iron limitation in the photoheterotrophic cells, but was abundant in phototrophic cells, irrespective of Fe-nutritional status. Like ferredoxin and cytochrome f, the non-Fe-containing PSII and PSI core proteins, D1 and PsaD, respectively, CB-839 molecular weight were also decreased 75% in photoheterotrophic iron-limited cells (0.1 μM Fe) but maintained in phototrophic iron-limited cells (Fig. 7). Fig. 7 Abundance of photosynthetic and respiratory proteins in photoheterotrophic versus phototrophic cells in response to iron nutrition. 20 μg of total protein was separated by denaturing polyacrylamide gel electrophoresis and immunoblotted for various photosynthetic and respiratory proteins. One of three representative experiments is shown Although photosynthesis requires more iron due to the high abundance of photosynthetic complexes in

the thylakoid membrane, the demand for iron per monomer is greater for respiration. Complex I requires the over most iron, containing a total of 8 iron–sulfur clusters (6 [Fe4S4] and 2 [Fe2S2]) for a total of 28 Fe atoms per complex I (Cardol et al. 2004; Sazanov 2007; Remacle et al. 2008). Complex II binds a total of 9 Fe atoms in the form of 3 iron–sulfur clusters (1 [Fe2S2], 1 [Fe3S4], and 1 [Fe4S4]) and 1 heme. Complex III contains 5 Fe atoms bound to 1 [Fe2S2] and 3 heme molecules, and complex IV utilizes 2 heme molecules to reduce oxygen to water. Since complex I contains the most iron, the abundance of iron-binding subunits of complex I was investigated. Surprisingly, similar to photosynthetic proteins, complex I subunits Nuo6 (Fe/S-binding) and Nuo7 (non-Fe/S-binding) were maintained in iron-limited (0.1-μM Fe) phototrophic cells, but decreased approximately 2-fold in heterophototrophic iron-limited cells, even though iron-limited heterophototrophic cells had a higher rate of oxygen consumption (Fig. 7; Table 2). Fe/S-binding Nuo8 was also more abundant in phototrophic when compared to photoheterotrophic cells (Fig. 7).