scholarly journals The production of amino acids by cell fractions, particularly rat-liver mitochondria

1963 ◽  
Vol 87 (1) ◽  
pp. 104-114 ◽  
Author(s):  
KGMM ALBERTI ◽  
W BARTLEY
Keyword(s):  
Amino Acids ◽  
Rat Liver ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Cell Fractions

scholarly journals The localization of proteolytic activity in rat liver mitochondria and its relation to mitochondrial swelling and aging

1969 ◽  
Vol 111 (5) ◽  
pp. 763-776 ◽  
Author(s):  
K. G. M. M. Alberti ◽  
W. Bartley
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Proteolytic Activity ◽  
Rat Liver ◽  
Acid Production ◽  
Liver Mitochondria ◽  
Ground Substance ◽  
Rat Liver Mitochondria ◽  
Amino Acid Production ◽  
Freezing And Thawing

1. On storage of rat liver mitochondria at 0°, water content, total amino acid content and leakage of protein all rose steadily over a 72hr. period. The initial ratio of intramitochondrial to extramitochondrial amino acid concentration lay between 18 and 24. Initially this rose, but it then fell to 1·9 at the end of storage. The concentration gradient between internal and external amino acids was relatively constant throughout the period. These processes were accentuated at 22° and 40°, the concentration gradient reaching 70μmoles/ml., water content rising to 8·3mg./mg. dry wt. and protein leakage reaching 42% of total mitochondrial protein. ‘Swelling agents’ produced no correlated changes in amino acid production and swelling. 2. Added glutamate was not concentrated within the pellet of whole or disrupted mitochondria. Endogenous amino acids were distributed evenly between the pellet and the supernatant of disrupted mitochondria. It is concluded that amino acids are produced within mitochondria and that adsorption and uptake from the medium do not contribute significantly to amino acids in the pellet. 3. β-Glycerophosphate, a lysosome protectant, increased amino acid production by rat liver mitochondria. Treatment with Triton X-100 and disruption by freezing and thawing showed that 56% of proteolytic activity was ‘free’ in whole mitochondria, whereas only 11% of acid phosphatase activity, a lysosomal enzyme, was ‘free’. 4. ‘Light’ mitochondria contained 30% more neutral proteolytic activity but 300% more acid phosphatase activity than ‘heavy’ mitochondria. 5. Electron micrographs of mitochondrial preparations showed less than one particle in 500 that could be identified as a lysosome. Treatment with Triton X-100 disrupted the structure of roughly 50% of the mitochondria; the rest appeared to retain their membrane, cristae and ground substance. Freezing and thawing caused gross swelling and loss of ground substance and rupture of external membranes. 6. Of the recovered proteolytic activity, 81% at pH7·4 and 70% at pH5·8 were found in the high-speed supernatant of broken mitochondria. A further fivefold increase in specific activity was found in the first protein fraction obtained by Sephadex G-50 gel filtration. 7. Between 60 and 80% of proteolytic activity was found in the 40–60%-saturated ammonium sulphate precipitate. Almost all of the soluble-fraction proteolytic activity could be recovered in a pH5·0 supernatant. 8. The results give no support to the view that mitochondrial neutral proteolytic activity reflects lysosomal content. 9. The possible role of intramitochondrial amino acid production and the proteolysis of internal barriers in passive swelling of mitochondria is discussed.

Transamination of Ring-Substituted L-Phenylalanines by an Extract from Rat Liver Mitochondria

1973 ◽  
Vol 51 (4) ◽  
pp. 407-411 ◽  
Author(s):  
J. H. Tong ◽  
B. A. Stoochnoff ◽  
A. D'Iorio ◽  
N. Leo Benoiton
Keyword(s):  
Amino Acids ◽  
Rat Liver ◽  
Liver Mitochondria ◽  
Tyrosine Aminotransferase ◽  
Rat Liver Mitochondria ◽  
Amino Group ◽  
Mitochondrial Enzyme ◽  
Soluble Enzyme ◽  
Keto Acids ◽  
Mitochondrial Extract

The L- and D-isomers of m-tyrosine, o-tyrosine, p-chlorophenylalanine (p-CP), and p-fluorophenylalanine (p-FP) were tested as substrates for the soluble tyrosine aminotransferase and a mitochondrial extract of rat liver by measuring the amino acids formed with 2-oxoglutarate, oxaloacetate, and pyruvate as acceptors. None of the above were substrates for the soluble enzyme. L-m-Tyrosine, L-p-CP, and L-p-FP were transaminated at substantial rates (16–25% of the rate for L-tyrosine) by the mitochondrial enzyme with all three keto acids as amino group acceptors. A slow but definite transamination of L-o-tyrosine by the mitochondrial enzyme was demonstrated using labeled 2-oxoglutarate as acceptor.

Effects of 5-5'-Diphenyl-2-thiohydantoin (DPTH) and Thyroxine on the Ultrastructure and Chemical Composition of Rat Liver

1971 ◽  
Vol 29 ◽  
pp. 570-571
Author(s):  
E. A. Elfont ◽  
R. B. Tobin ◽  
D. G. Colton ◽  
M. A. Mehlman
Keyword(s):  
Chemical Composition ◽  
Rat Liver ◽  
Future Study ◽  
Mode Of Action ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Effective Inhibitor ◽  
Biochemical Effects ◽  
Glycerophosphate Dehydrogenase ◽  
Stimulation Of

Summary5,-5'-diphenyl-2-thiohydantoin (DPTH) is an effective inhibitor of thyroxine (T4) stimulation of α-glycerophosphate dehydrogenase in rat liver mitochondria. Because this finding indicated a possible tool for future study of the mode of action of thyroxine, the ultrastructural and biochemical effects of DPTH and/or thyroxine on rat liver mere investigated.Rats were fed either standard or DPTH (0.06%) diet for 30 days before T4 (250 ug/kg/day) was injected. Injection of T4 occurred daily for 10 days prior to sacrifice. After removal of the liver and kidneys, part of the tissue was frozen at -50°C for later biocheailcal analyses, while the rest was prefixed in buffered 3.5X glutaraldehyde (390 mOs) and post-fixed in buffered 1Z OsO4 (376 mOs). Tissues were embedded in Araldlte 502 and the sections examined in a Zeiss EM 9S.Hepatocytes from hyperthyroid rats (Fig. 2) demonstrated enlarged and more numerous mitochondria than those of controls (Fig. 1). Glycogen was almost totally absent from the cytoplasm of the T4-treated rats.

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