Beef heart

FIGURE 21.9 Typical visible absorption spectra of cytochromes, (a) Cytochrome c, reduced spectrum (b) cytochrome c, oxidized spectrum (c) the difference spectrum (a) minus (b) (d) beef heart mitochondrial particles room temperature difference (reduced minus oxidized) spectrum (e) beef heart submitochondrial particles same as (d) but at 77 K. a- and /3- bauds are labeled, and in (d) and (e) the bauds for cytochromes a, h and c are indicated. 

Fig. 6. Comparison of VTMCD spectra for biological [Fe3S4] clusters. (A) D. gigas Fdll (20) (B) P. furiosus 3Fe Fd (42) (C) A. vinelandii Fdl (70) (D) T. thermophilus 7Fe Fd (70) (E) E. coli nitrate reductase (24) (F) E. coli fumarate reductase (53) (G) spinach glutEimate synthase (25) (H) beef heart aconitase (27). Spectra were recorded at temperatures between 1.5 and 70 K with an apphed magnetic field of 4.5 T (sdl trEmsitions increase in intensity with decreasing temperature). BEmds originating from minor heme contaminEmts Eire indicated by an asterisk.

Vibrational Frequencies (cm" ), Isotope Downshifts (cm" in Parentheses), and Assignments for the Fe-S Stretching Modes of Oxidized FeaSJSa Centers in Beef Heart Aconitase, a. vinelandii Fdl, T. thermophilus Fd, P. furiosus Fd, D. gigas Fdll, and... 

Fig. 5. Structural comparison of the water-soluble fragments of the Rieske proteins from (a) spinach chloroplasts and (b) beef heart mitochondria. Conserved smd vEiriable regions are highlighted and the conserved /3-loop discussed in Fig. 6 is denoted by darker gray on the rear of the molecules.

Fig. 6. Sequence comparisons of Rieske proteins from spinach chloroplasts, beef heart mitochondria, green sulfur bacteria, and firmicutes. The extended insertion of proteobacterial Rieske proteins as compared to the mitochondrial one is indicated by a dotted arrow. The redox-potential-influencing Ser residue is marked by a vertical arrow. The top and the bottom sequence numberings refer to the spinach and bovine proteins, respectively. Fully conserved residues are marked by dark shading, whereas the residues conserved in the b6f-group are denoted by lighter shading.

Ethanol and choline glycerolipids were isolated from calf brain and beef heart lipids by PTLC using silica gel H plates. Pure ethanol amine and choline plasmalogens were obtained with a yield of 80% [74]. Four phosphohpid components in the purple membrane (Bacteriorhodopsin) of Halobacterium halobium were isolated and identified by PTLC. Separated phosphohpids were add-hydrolyzed and further analyzed by GC. Silica gel G pates were used to fractionate alkylglycerol according to the number of carbon atoms in the aliphatic moiety [24]. Sterol esters, wax esters, free sterols, and polar lipids in dogskin hpids were separated by PTLC. The fatty acid composition of each group was determined by GC. 

Ghose, S. and Mattiasson, B., Evaluation of displacement chromatography for the recovery of lactate dehydrogenase from beef heart under scale-up conditions,. Chromatogr., 547, 145, 1991. 

The Wassermann substance17 is prepared by extracting various animal organs, particularly beef heart, with alcohol, and its lipidal nature was early recognized. Pangborn174 described the preparation of a new phospholipid termed cardiolipin from beef heart and claimed that it was the essential constituent of the Wassermann substance. On hydrolysis it gave a fatty acid and a phosphorylated polysaccharide. In a later communication17 however, the carbohydrate constituent is stated... 

Baird, B.A., and Hammes, G.G. (1977) Chemical cross-linking studies of beef heart mitochondrial coupling factor 1./. Biol. Chem. 252, 4743-4748. 

LOX catalyzed the oxidation of arachidonoylphosphatidylcholine at both carbon-12 and carbon-15. Later on, it has been found [21] that reticulocyte lipoxygenase oxidized rat liver mitochondrial membranes, beef heart submitochondrial particles, rat liver endoplasmic membranes, and erythrocyte plasma membranes without preliminary release of unsaturated acids by phospholipases. 

Schnurr et al. [22] showed that rabbit 15-LOX oxidized beef heart submitochondrial particles to form phospholipid-bound hydroperoxy- and keto-polyenoic fatty acids and induced the oxidative modification of membrane proteins. It was also found that the total oxygen uptake significantly exceeded the formation of oxygenated polyenoic acids supposedly due to the formation of hydroxyl radicals by the reaction of ubiquinone with lipid 15-LOX-derived hydroperoxides. However, it is impossible to agree with this proposal because it is known for a long time [23] that quinones cannot catalyze the formation of hydroxyl radicals by the Fenton reaction. Oxidation of intracellular unsaturated acids (for example, linoleic and arachidonic acids) by lipoxygenases can be suppressed by fatty acid binding proteins [24]. 

This mechanism is now considered to be of importance for the protection of LDL against oxidation stress, Chapter 25.) The antioxidant effect of ubiquinones on lipid peroxidation was first shown in 1980 [237]. In 1987 Solaini et al. [238] showed that the depletion of beef heart mitochondria from ubiquinone enhanced the iron adriamycin-initiated lipid peroxidation whereas the reincorporation of ubiquinone in mitochondria depressed lipid peroxidation. It was concluded that ubiquinone is able to protect mitochondria against the prooxidant effect of adriamycin. Inhibition of in vitro and in vivo liposomal, microsomal, and mitochondrial lipid peroxidation has also been shown in studies by Beyer [239] and Frei et al. [240]. Later on, it was suggested that ubihydroquinones inhibit lipid peroxidation only in cooperation with vitamin E [241]. However, simultaneous presence of ubihydroquinone and vitamin E apparently is not always necessary [242], although the synergistic interaction of these antioxidants may take place (see below). It has been shown that the enzymatic reduction of ubiquinones to ubihydroquinones is catalyzed by NADH-dependent plasma membrane reductase and NADPH-dependent cytosolic ubiquinone reductase [243,244]. 

Figure 6.3 (a) Visible absorption spectrum of cytochrome c in its reduced and oxidized states, (b) The three separate a bands in the visible spectrum of beef heart mitochondria (below) indicating the presence of cytochromes a, b and c, with the spectrum of cytochrome c (above) as reference. (From Voet and Voet, 2004. Reproduced with permission from John Wiley Sons., Inc.)... 

Figure 14.9 Schematic representation of the redox metals of beef heart CcOX with their relative distances. (From Brunori et al., 2005. Copyright 2005, with permission from Elsevier.)...

The mitochondrial respiratory parameters have also been employed to determine the toxicity of surfactants, including anionic (LAS), nonionic (NPEO) and their metabolites, sulfophenyl carboxylates (SPCs), NP and nonylphenoxy carboxylate (NPECi) [37]. The system employed was the in vitro response of submitochondrial particles from beef heart. The EC50 toxicity calculated as the reduction rate of NAD+ ranged from 0.61 mg L-1 for a commercial LAS mixture to 18 000 mg L-1 for SPCs, and 1.3 mg L-1, 8.2 and 1.8mgL 1 for NPEOio, NPECi and NP, respectively. These results indicate that from the toxicity perspective, LAS is the compound demanding increased attention, while for NPEO, the parental compound and the metabolites must be quantified. 

By 1949 low temperature spectroscopy had been introduced. With this technique Keilin and Hartree detected a further component in the electron transfer chain which had a sharp band at 552 nm. They later showed it to be identical with cytochrome cj, which had first been observed by Yakushiji and Okunuki (1940) during succinate oxidation by cyanide-inhibited beef heart muscle. As the oxidation of cytochrome C was accelerated by cytochrome c, Okunuki and Yakushiji (1941) had placed C] in the chain in the order... 

ATP- Pi exchange Beef heart Rat liver — 27 27 40 42 Penniston (1971) Penniston (1971)... 

Beyer, K., KUngenberg, M., 1985, ADP/ATP carrier protein from beef heart mitochondria has high amounts oftightly bound cardioUpin, as revealed by P nuclear magnetic resonance. Biochemistry 24 3821-3826. 

Beyer, K., and Nuscher, B., 1996, Specific cardiolipin binding interferes with labeling of sulfhydryl residues in the adenosine diphosphate/adenosine triphosphate carrier protein from beef heart mitochondria, Biochemistry 35 15784-15790. 

A newly emerging class of iron sulfur proteins are those with 3Fe-XS centers. Aerobically isolated, inactive beef heart aconitase has a 3Fe-4S center which has... 

Figure 1. EPR absorption derivative spectrum of aconitase (as isolated from beef heart mitochondria) in 100 mM potassium phosphate, pH 7.0. Experimental conditions for obtaining the EPR spectrum were 10 K, 100 microwatts power, 0.8 milliTesla (mT) modulation amplitude, and 9.42 GHz microwave frequency.

Figure 2. Zero-field MSssbauer spectra of dithionite reduced three-iron clusters of (A) D. gigas ferredoxin n, (B) A. vinelandii ferredoxin I, and (C) aconitase isolated from beef heart mitochondria. (Reproduced with permission fixim Ref. 38. Copyright 1982 Elsevier.)...

Bieber LL, Choi YR (1977) Isolation and identification of aliphatic short-chain acylcarnitines from beef heart possible role for carnitine in branched-chain amino acid metabolism. Proc Natl Acad Sci U S A 74 2795-2798... 

Hall, N. Addis, P. DeLuca, M. Mitochondrial creatine kinase. Physical and kinetic properties of the purified enzyme from beef heart. Biochemistry, 18, 1745-1751 (1979)... 

Colomb, M.G. Cheruy, A. Vignais, P.V. Nucleoside diphosphokinase from beef heart cytosol. I. Physical and kinetic properties. Biochemistry, 11, 3370-3378 (1972)... 

Albrecht, G.J. Purification and properties of nucleoside triphosphate-adenosine monophosphate transphosphorylase from beef heart mitochondria. Biochemistry, 9, 2462-2470 (1970)... 

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