Influence of charge and polarity on the redox potentials of high-potential iron-sulfur proteins: evidence for the existence of two groups.
We have investigated the HiPIPs from Ectothiorhodospira vacuolata (iso-1 and iso-2), Chromatium vinosum, Rhodocyclus gelatinosus, Rhodocyclus tenuis (strain 2761), Rhodopila globiformis, and Rhodospirillum salinarum (iso-2) by direct electrochemistry. Using a glassy carbon electrode with a negatively charged surface, direct, unpromoted electrochemistry is possible with the positively charged HiPIPs. With the negatively charged HiPIPs, the positively charged and flexible bridging promoter poly(L-lysine) is required. The stability of the response can be improved by morpholin, aspartate, tryptophan, or 4,4'-dipyridyl. These "stabilizers" prevent the blocking of the electrode by denatured protein. The redox potential of 500 mV found for R. salinarum iso-2 is the highest HiPIP potential reported. The presence of histidines in the sequence does not per se predict a pH-dependent redox potential. Only C. vinosum and R. gelatinosus HiPIPs show a weak but significant pH dependence with a difference of 35 mV between the low- and the high-pH form and maximum slopes of -20 mV/unit. The dependence of the midpoint potential on temperature and on ionic strength varies over the different HiPIPs. The dependence of the potentials on square root of I cannot be fully explained by the Debye-Hückel theory because the linearity exceeds the limiting concentration and only small negative slopes are observed (o to -28 mV/square root of M) Combination of the sequences, the optical spectra, the overall charges, and the redox thermodynamics suggests that existence of two groups of HiPIPs. One group consists of Chromatium-like HiPIPs with redox potentials between 300 and 350 mV, modulated only by the solvation of the cluster. The second group is formed by Ectothiorhodospira-like HiPIPS with potentials between 50 and 500 mV, modulated by the overall charge of the peptide (25 mV/unit) and by the solvation of the cluster.
Heering
H A
HA
Department of Biochemistry, Agricultural University Wageningen, The Netherlands.
Bulsink
B M
BM
Hagen
W R
WR
Meyer
T E
TE
D000596:Amino Acids / Q000494:pharmacology; D001419:Bacteria / Q000737:chemistry*; D004315:Doxapram / Q000031:analogs & derivatives / Q000494:pharmacology; D004563:Electrochemistry; D004579:Electron Transport; D006863:Hydrogen-Ion Concentration; D007506:Iron-Sulfur Proteins / Q000737:chemistry* / Q000378:metabolism; D007526:Isoelectric Point; D009994:Osmolar Concentration; D010084:Oxidation-Reduction; D011107:Polylysine / Q000378:metabolism; D013057:Spectrum Analysis; D013696:Temperature; D013816:Thermodynamics
D000596:Amino Acids; D007506:Iron-Sulfur Proteins; D011107:Polylysine; C029505:2-ketodoxapram; D004315:Doxapram