Absorption of antisera for studies on specific enzyme turnover

Walker, John H.; Betts, S A; Manning, Ray; Mayer, R. John

doi:10.1042/bj1590355

Cited by 17 publications

(10 citation statements)

References 26 publications

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“…Monospecific antiserum to acetyl-CoA carboxylase was prepared as described by Walker et al (1976) and was a gift from Dr. R. J. Mayer (Department of Biochemistry, University Hospital and Medical School, Nottingham, U.K.). Adrenaline was obtained as the bitartrate salt from BDH (Poole, Dorset, U.K.).…”

Section: Experimental Chemicals and Materialsmentioning

confidence: 99%

Adrenaline and the regulation of acetyl-coenzyme A carboxylase in rat epididymal adipose tissue. Inactivation of the enzyme is associated with phosphorylation and can be reversed on dephosphorylation

Brownsey¹,

Hughes²,

Denton³

1979

Biochemical Journal

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Section: Experimental Chemicals and Materialsmentioning

confidence: 99%

Adrenaline and the regulation of acetyl-coenzyme A carboxylase in rat epididymal adipose tissue. Inactivation of the enzyme is associated with phosphorylation and can be reversed on dephosphorylation

Brownsey¹,

Hughes²,

Denton³

1979

Biochemical Journal

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“…Assuming similar antigen to antibody ratios in immunoprecipitates, comparable amounts of proteins were precipitated from mitochondria, microsomes and cytosol (Table 1). Immunoprecipitates derived from interaction with a rat liver particle-free supernatant and antibodies to rat liver cytochrome oxidase were also described by Walker et al [67]. Immunological relationship to cytochrome oxidase, however, could not be demonstrated.…”

Section: Discussionmentioning

confidence: 99%

Immunoprecipitation of a Cytoplasmic Precursor of Rat-Liver Cytochrome Oxidase

1978

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A simple method for the isolation of rat liver cells is described. The cells are shown, by an isotope dilution method, to maintain a constant rate of protein synthesis for 8 h of incubation. Antibodies to purified rat liver cytochrome oxidase were raised in rabbits and used to investigate the labeling of cytochrome oxidase in isolated rat liver cells and in vivo. The data demonstrate the occurrence of a precursor of the subunits of cytochrome oxidase that are synthesized in the cytoplasm.1. Dodecylsulfate gel electrophoresis of the immunoprecipitates from isolated rat liver cells that had been labeled with [35S]methionine for 1 h showed a single radioactive peak with a molecular weight of 50000.2. Judged by the effects of cycloheximide and chloramphenicol the labeled protein is synthesized on cytoplasmic ribosomes.3. After labeling for 1 h in vivo with [3H]leucine the labeled protein appears to be exclusively associated with the hepatic niicrosomal fraction.4. Ouchterlony double-diffusion analysis demonstrated immunological relationship between the precipitates from microsomes and cytochrome oxidase. In addition to the precipitates derived from mitochondria and microsomes immunoprecipitates were also obtained from the cytosol in comparable amounts ; these again were immunologically related.The occurrence of large amounts of precursor(s) (or degradation products) of cytochrome oxidase in rat liver fractions is interpreted in terms of a regulatory pool for amino acid homeostasis in the organism.Cytochrome oxidase, the terminal oxygen acceptor of the respiratory chain in the inner mitochondrial membrane, represents a hydrophobic enzyme complex that is composed of seven protein subunits, two heme a groups, two copper atoms and tightly bound lipids (for reviews see [l-31). The biosynthesis of the enzyme is a complex process involving the assembly of four small subunits that are coded for in the nucleus and synthesized on cytoplasmic ribosomes, and three larger subunits that are coded for and synthesized within the mitochondria (for reviews see . Although these results are mainly based on studies with yeast similar data on subunit composition [9, of the cytoplasmically synthesized subunits and on the mechanism of assembling the enzyme complex. Poyton et al. described the extramitochondrial occurrence of a precursor protein for the four small subunits of yeast cytochrome oxidase which stimulates the synthesis of the three large subunits in mitochondria and which is immunoprecipitated from a postpolysomal supernatant fraction by antibodies to the holoenzyme 129,301. This protein has a molecular weight of 55000 and is thought to contain a small peptide in addition to the sequences of the four cytoplasmically synthesized subunits of cytochrome oxidase. Rucker et al. also presented evidence for the occurrence of precursor subunits VI and VII in the postribosomal cytoplasmic supernatant fraction of Neurospora crassa cells [31]. An intermediate in the assembly of mitochondria1 and cytoplasmically synthesized subunits of cy...

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“…monoamine oxidase) (Walker et al, 1976;Mayer & Walker, 1978). Highly purified liver monoamine oxidase was used to raise the antiserum to minimize the possibility of producing antibodies to contaminating antigens (Russell et al, 1979).…”

Section: Discussionmentioning

confidence: 99%

Immunochemical characterization of monoamine oxidase from human liver, placenta, platelets and brain cortex

Russell¹,

Davey²,

Mayer³

1979

Biochemical Journal

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1. Antiserum raised to purified human liver monoamine oxidase was used to characterize the monoamine oxidase from human liver, brain cortex, placenta and platelets. 2. Antibodies to monoamine oxidase were purified by adsorption with a mitochondrial preparation. 3. Monoamine oxidase was present in liver particle-free supernatant as measured by enzyme activity and immunodiffusion. 4. Multiple precipitin lines were obtained on immunodiffusion analysis against the purified liver enzyme. It is proposed that this is due to either aggregation or to differential lipid binding. 5. The results suggest that the functionally different enzymes found in liver, brain cortex, platelets and placenta are immunochemically related and may be identical.

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Absorption of antisera for studies on specific enzyme turnover

Cited by 17 publications

References 26 publications

Adrenaline and the regulation of acetyl-coenzyme A carboxylase in rat epididymal adipose tissue. Inactivation of the enzyme is associated with phosphorylation and can be reversed on dephosphorylation

Adrenaline and the regulation of acetyl-coenzyme A carboxylase in rat epididymal adipose tissue. Inactivation of the enzyme is associated with phosphorylation and can be reversed on dephosphorylation

Immunoprecipitation of a Cytoplasmic Precursor of Rat-Liver Cytochrome Oxidase

Immunochemical characterization of monoamine oxidase from human liver, placenta, platelets and brain cortex

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