title: The Multicenter Aerobic Iron Respiratory Chain of Acidithiobacillus ferrooxidans Functions as an Ensemble with a Single Macroscopic Rate Constant.
abstract: Electron transfer reactions among three prominent colored proteins in intact cells of Acidithiobacillus ferrooxidans were monitored using an integrating cavity absorption meter that permitted the acquisition of accurate absorbance data in suspensions of cells that scattered light. The concentrations of proteins in the periplasmic space were estimated to be 350 and 25 mg/ml for rusticyanin and cytochrome c, respectively; cytochrome a was present as one molecule for every 91 nm(2) in the cytoplasmic membrane. All three proteins were rapidly reduced to the same relative extent when suspensions of live bacteria were mixed with different concentrations of ferrous ions at pH 1.5. The subsequent molecular oxygen-dependent oxidation of the multicenter respiratory chain occurred with a single macroscopic rate constant, regardless of the proteins' in vitro redox potentials or their putative positions in the aerobic iron respiratory chain. The crowded electron transport proteins in the periplasm of the organism constituted an electron conductive medium where the network of protein interactions functioned in a concerted fashion as a single ensemble with a standard reduction potential of 650 mV. The appearance of product ferric ions was correlated with the reduction levels of the periplasmic electron transfer proteins; the limiting first-order catalytic rate constant for aerobic respiration on iron was 7,400 s(-1). The ability to conduct direct spectrophotometric studies under noninvasive physiological conditions represents a new and powerful approach to examine the extent and rates of biological events in situ without disrupting the complexity of the live cellular environment.
authors.affiliation: From the College of Pharmacy, Xavier University of Louisiana, New Orleans, Louisiana 70125.
authors.forename: Ting-Feng
authors.initials: TF
authors.lastname: Li
authors.affiliation: From the College of Pharmacy, Xavier University of Louisiana, New Orleans, Louisiana 70125.
authors.forename: Richard G
authors.initials: RG
authors.lastname: Painter
authors.affiliation: From the College of Pharmacy, Xavier University of Louisiana, New Orleans, Louisiana 70125.
authors.forename: Bhupal
authors.initials: B
authors.lastname: Ban
authors.affiliation: From the College of Pharmacy, Xavier University of Louisiana, New Orleans, Louisiana 70125 rblake@xula.edu.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Blake
mesh_terms: D042763:Acidithiobacillus / Q000737:chemistry / Q000378:metabolism*; D000332:Aerobiosis; D045264:Cytochromes a / Q000378:metabolism; D045304:Cytochromes c / Q000378:metabolism; D004579:Electron Transport; D007501:Iron / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D010084:Oxidation-Reduction
chemical_list: D045304:Cytochromes c; D045264:Cytochromes a; D007501:Iron
keywords: bacteria; cytochrome; electron transport; integrating cavity absorption meter; macromolecular crowding; oxidation-reduction (redox); respiratory chain