Assembly of the Mitochondrial Membrane System

Rubin, Meryl S.; Tzagoloff, Alexander

doi:10.1016/s0021-9258(19)43767-8

Cited by 136 publications

(11 citation statements)

References 24 publications

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“…On the other hand, the polypeptide might well retain its native conformation to a greater extent; the polypeptide would probably not assume the flexible extended rod found for water soluble polypeptides interacting with NaDodS04 (Tanford, 1973), so it would pass through the gel more quickly. Other integral membrane proteins have been shown to run at anomalously low molecular weights on NaDodS04-PAGE (Bridgen and Walker, 1976;Simons and Kááriáinen, 1970;Rubin and Tzagoloff, 1973;Strittmatter, 1971, 1973;Kyte, 1972). These results suggest that one should consider the possibility of an anomalously low molecular weight determined by NaDodS04-PAGE for an integral membrane polypeptide.…”

Section: Discussionmentioning

confidence: 90%

Isolation and characterization of an organic solvent soluble polypeptide component from photoreceptor complexes of Rhodospirillum rubrum

Tonn¹,

Gogel²,

Loach

1977

Biochemistry

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An organic solvent soluble polypeptide has been isolated from photoreceptor complexes and chromatophores of Rhodospirillum rubrum. After extraction of the protein from lyophilized samples with 1:1 chloroform-methanol, it was purified by column chromatography. Its isoelectric point determined by isoelectric focusing was 7.10. When analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the purified polypeptide ran as a single band of an apparent molecular weight of 12 000. However, according to amino acid analysis, the minimal molecular weight based on one histidine residue per polypeptide is 19 000. The polypeptide contains no cysteine and no tyrosine. Amino acid analysis indicated that three methionines were present per histidine residue and cyanogen bromide cleavage gave four smaller peptides which were isolated by two-dimensional electrophoresis and chromatography. Spectroscopic analysis indicated the presence of three tryptophan residues per histidine and N-bromosuccinamide cleavage also gave four smaller peptides which could be isolated by two-dimensional electrophoresis and chromatography. The C-terminal amino acid was shown to be glycine by two methods, while the N-terminal amino acid appears to be blocked. The organic solvent soluble polypeptide accounts for approximately 50% of the chromatophore protein and seems to bind the antenna bacteriochlorophyll and carotenoid molecules. Using this procedure, organic solvent soluble polypeptides were isolated from several photosynthetic bacteria and were found to have substantially different amino acid contents.

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Section: Discussionmentioning

confidence: 90%

Isolation and characterization of an organic solvent soluble polypeptide component from photoreceptor complexes of Rhodospirillum rubrum

Tonn¹,

Gogel²,

Loach

1977

Biochemistry

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“…Nonionic surfactants were, in fact, regarded as the most mild and suitable surface-active materials for preparing lipid-free membrane proteins in their native conformation . However, it was also found that nonyl-phenol ethoxylates could cause the loss of biological activities in some globular proteins, e.g., cytochrome oxidase, glucose-6-phosphatase, and adenylate cyclase . Tanford et al , later showed that nonionic surfactants interact with globular proteins and that their binding to native BSA was specific to the hydrophobic domains, in contrast to the cooperative association characteristic of anionic surfactants, as described below.…”

Section: Resultsmentioning

confidence: 99%

Binding of Surfactants onto Preadsorbed Layers of Bovine Serum Albumin at the Silica−Water Interface

Thomas³

1998

J. Phys. Chem. B

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The binding of nonionic and anionic surfactants from bulk solutions onto preadsorbed layers of bovine serum albumin (BSA) at the hydrophilic silicon oxide/water interface has been studied using specular neutron reflection. The distributions of surfactant, protein, and water at the solid/aqueous interface were separately determined by using deuterium labeling of the surfactant and water. The preadsorbed BSA layers were formed by equilibrating the hydrophilic oxide surface with a 0.15 g dm -3 BSA solution, which produced a densely packed uniform BSA layer of thickness 35 ( 3 Å. The possible association of nonionic surfactant pentaethylene glycol monododecyl ether (C 12 E 5 ) with BSA at the interface was examined by following the change in neutron reflectivity before and after surfactant addition. That no measurable difference between these reflectivity profiles was observed suggests that the nonionic surfactant did not bind to the adsorbed BSA. The binding of sodium dodecyl sulfate (SDS) onto the BSA layer was studied over a wide SDS concentration range. Noticeable binding was detected at an SDS concentration of 1 × 10 -4 M, and as the SDS concentration increases the thickness of the mixed layer also increases. At [SDS] ) 1 × 10 -3 M the reflectivity profile from the mixed layer became identical to that between the bare oxide/D 2 O interface, suggesting that at this SDS concentration the mixture is completely removed from the interface. The surface excesses of BSA and SDS, and the structural distributions of BSA, SDS and water at each SDS concentration were obtained by simultaneously fitting a single structural model to the set of measured neutron reflectivity profiles. The results show that at low SDS concentrations the structural profiles of the interfacial components are well approximated as uniform layer distributions and as SDS concentration is increased the distributions become unsymmetrical. The binding of SDS results in an expansion of the preadsorbed BSA layer from 35 ( 3 Å in the absence of SDS to some 80 Å at 3 × 10 -4 M, suggesting considerable structural deformation of the protein. The volume ratio of SDS to BSA in the mixed layer was found to be 0.45, in close agreement with the literature value for the binding of SDS onto denatured protein in the bulk, suggesting that the protein in the adsorbed complex is also denatured.

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“…Much of the variability in results has been the consequence of differences in isolation procedures or conditions of gel electrophoresis, failure to adequately dissociate the enzyme, or selective aggregation of subunits during preparation of the samples for gel electrophoresis, and the presence of contaminating polypeptides. There is general agreement that yeast and Neurospora enzymes can be resolved into seven polypeptides, each present in one copy (Rubin & Tzagoloff, 1973a;Sebald et al, 1973;Poyton & Schatz, 1975;Phan & Mahler, 1976a), with similar molecular weights of the individual subunits in the two organisms. The enzyme purified from animal cells has been resolved into six to eight subunits (Weiss et al, 1972;Koch, 1976;Rascati & Parsons, 1979a;Hochli & Hackenbrock, 1978;Rubin & Tzagoloff, 1973a;Phan & Mahler, 1976a;Yatscoff et al, 1977;Capaldi & Hayashi, 1972;Kuboyama et al, 1972;Capaldi et al, 1977;Buse & Steffens, 1976).…”

Section: Discussionmentioning

confidence: 98%

Isolation, subunit composition, and site of synthesis of human cytochrome c oxidase

Hare¹,

Ching²,

Attardi³

1980

Biochemistry

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Cytochrome c oxidase (ferrocytochrome c:oxygen oxidoreductase, EC 1.9.3.1), the terminal oxidase of the respiratory chain in eucaryotic cells, has been purified from human placenta mitochondria. Seven polypeptides have been identified reproducibly by high-resolution electrophoresis of the enzyme complex through sodium dodecyl sulfate (Na-DodSO4)--urea polyacrylamide gels; these correspond closely in size to the subunits of beef heart cytochrome c oxidase. When HeLa cells, grown in suspension culture, were pulse-labeled with [35S]methionine in the presence of cycloheximide to inhibit cytoplasmic protein synthesis and chased with an excess of unlabeled methionine in the absence of the drug, the mitochondrially synthesized polypeptides were resolved into at least 17 components by NaDodSO4--urea polyacrylamide gel electrophoresis. After labeled HeLa mitochondria were mixed with human placenta mitochondria and the cytochrome c oxidase was isolated, three of the labeled components were found to copurify with the three largest subunits of the complex. We conclude that human cytochrome c oxidase contains seven subunits, the three largest of which are synthesized on mitochondrial ribosomes, while the other four are synthesized in the cytoplasm.

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Assembly of the Mitochondrial Membrane System

Cited by 136 publications

References 24 publications

Isolation and characterization of an organic solvent soluble polypeptide component from photoreceptor complexes of Rhodospirillum rubrum

Isolation and characterization of an organic solvent soluble polypeptide component from photoreceptor complexes of Rhodospirillum rubrum

Binding of Surfactants onto Preadsorbed Layers of Bovine Serum Albumin at the Silica−Water Interface

Isolation, subunit composition, and site of synthesis of human cytochrome c oxidase

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