https://www.jbc.org/content/246/17/5477.full.pdf

Partial Resolution of the Enzymes Catalyzing Oxidative Phosphorylation XXV. RECONSTITUTION OF VESICLES CATALYZING 32Pi—ADENOSINE TRIPHOSPHATE EXCHANGE

A23187: a divalent cation ionophore.

It was obviously my hope that the chemiosmotic rationale of vectorial metabolism and biological energy transfer might one day come to be generally accepted, and I have done my best to argue in favour of that state of affairs for more than twenty years. But, it would have been much too presumptuous to have expected it to happen. Of course, I might have been wrong, and in any case, was it not the great Max Planck (1928, 1933) who remarked that a new scientific idea does not triumph by convincing its opponents, but rather because its opponents eventually die? The fact that what began as the chemiosmotic hypothesis has now been acclaimed as the chemiosmotic theory-at the physiological level, even if not at the biochemical level-has therefore aroused in me emotions of astonishment and delight in full and equal measure, which are all the more heartfelt because those who were formerly my most capable opponents are still in the prime of their scientific lives. I shall presently explain the difference between the physiological and the biochemical levels at which the chemiosmotic theory has helped to promote useful experimental research. But let me first say that my immediate and deepest impulse is to celebrate the fruition of the creative work and benevolent influence of the late David Keilin, one of the greatest of biochemists and-to me, at least-the kindest of men, whose marvellously simple studies of the cytochrome system, in animals, plants and microorganisms (Keilin, 1925), led to the original fundamental idea of aerobic energy metabolism: the concept of the respiratory chain (Keilin, 1929; and see Nicholls, 1963; King, 1966). Perhaps the most fruitful (and surprising) outcome of the development of the notion of chemiosmotic reactions is the experimental stimulus and guidance it has provided in work designed to answer the following three elementary questions about respiratory chain systems and analogous photoredox chain systems: What is it? What does it do? How does it do it? The genius of David Keilin led to the revelation of the importance of these questions. In this lecture, I hope to show that, as a result of the painstaking work of many biochemists, we can now answer the first two in general principle, and that considerable progress is being made in answering the third. Owing to the broad conceptual background, and the very wide range of practical application of the chemiosmotic theory, I have had …

https://science.sciencemag.org/content/sci/206/4423/1148.full.pdf

Keilin's respiratory chain concept and its chemiosmotic consequences.

The Synthesis and Ion Bindings of Synthetic Multidentate Macrocyclic Compounds.

The energy-linked reaction of calcium with mitochondria.

Publisher Summary This chapter describes the isolation of liver or kidney mitochondria. The selected tissue is disrupted by homogenization in cold isotonic sucrose. Differential centrifugation is then employed to separate the mitochondria from cell debris, red blood cells, nuclei, microsomes, and soluble components. For the isolation of liver mitochondria, the homogenate is distributed into Lusteroid centrifuge cups and centrifuged at 600 g for 10 minutes. The supernatant fraction is decanted and saved. The pellets may be dispersed by using the side of a stirring rod against the wall of the cup or by handoperating the homogenizer. The resuspended material is centrifuged at 600 g for 10 minutes. The supernatant fractions are combined. The pellets are discarded. This washing contributes not only to the yield of the final mitochondrial preparation, but also to its integrity, apparently by permitting the recovery of the larger mitochondria. For the isolation of kidney, the kidney capsule is removed by gently squeezing the kidney through the thumb and forefinger. The kidney is then cut sagittally. The medullary portion is removed and discarded. Mitochondria are then prepared from the cortex following the method described for liver, with the exception that the mitochondrial pellet need be washed only once. Method for testing the quality of mitochondria is also discusses in the chapter.

[15] Isolation of liver or kidney mitochondria

Antibiotic studies. ii. inhibition of phosphoryl transfer in mitochondria by oligomycin and aurovertin.

Antibiotics as tools for metabolic studies. V. Effect of nonactin, monactin, dinactin, and trinactin on oxidative phosphorylation and adenosine triphosphatase induction.

Antibiotics as Tools for Metabolic Studies. IV. Comparative Effectiveness of Oligomycins A, B, C, and Rutamycin as Inhibitors of Phosphoryl Transfer Reactions in Mitochondria*

Conformational features of the oligoribonucleic acid (oligo-RNA) A1-U2-C3-C4-A5 are explored by proton nuclear magnetic resonance (NMR). The sequence is a molecular cognate of a portion of the T psi C loop and stem regions of yeast tRNAPhe. The molecule forms at least two classes of flexible yet ordered structures. Class I states are similar in spectral properties to the component oligomers, AU, AUC, and AUCC, and are likely to be standard right-helical structures. Class II states are characterized by a 2'-endo pucker at A1 and unusually large shielding of several C3 and U2 protons. Most of these features are consistent with identifying the class II solution structures with the "arch" conformation for the T psi C region determined by X-ray crystallography of yeast tRNAPhe.

Diane Johnson

Papers

[15] Isolation of liver or kidney mitochondria

Antibiotic studies. ii. inhibition of phosphoryl transfer in mitochondria by oligomycin and aurovertin.

Antibiotics as tools for metabolic studies. V. Effect of nonactin, monactin, dinactin, and trinactin on oxidative phosphorylation and adenosine triphosphatase induction.

Antibiotics as Tools for Metabolic Studies. IV. Comparative Effectiveness of Oligomycins A, B, C, and Rutamycin as Inhibitors of Phosphoryl Transfer Reactions in Mitochondria*

Unusual structures in single-stranded ribonucleic acid: proton nuclear magnetic resonance of AUCCA in deuterium oxide.