scholarly journals Regulation of the breakdown rates of biotin-containing proteins in Swiss 3T3-L1 cells

1988 ◽  
Vol 251 (3) ◽  
pp. 749-755 ◽  
Author(s):  
C S Chandler ◽  
F J Ballard

1. Degradation rate constants for individual biotin-labelled proteins were measured in Swiss 3T3-L1 adipocytes that had been incubated with inhibitors of autophagy or of lysosomal proteolysis. 2. Inhibitory effects produced by 10 mM-3-methyladenine and a combination of 5 mM-NH4Cl and leupeptin (50 micrograms/ml) were approximately equal. The inclusion of NH4Cl did not significantly enhance the responses to 3-methyladenine, suggesting that autophagy was already maximally inhibited. 3. The extent of inhibition by 3-methyladenine or by the NH4Cl/leupeptin mixture was similar for the cytosolic enzyme acetyl-CoA carboxylase and for the three mitochondrial carboxylases. This inhibition averaged 50%. The breakdown rate of a more-stable 38 kDa biotin-containing mitochondrial protein was more responsive to the inhibitory agents. These results are best explained by mitochondrial proteolysis occurring via a combination of the degradation of whole mitochondria within autophagic vacuoles, supplemented by the selective intramitochondrial breakdown of more labile proteins. 4. A number of intermediate products in the degradation of biotin-containing proteins were detected. Differences in the patterns of radioactivity between these peptides after incubation of cells in the presence of inhibitors of the breakdown process provided evidence that some peptides were produced before autophagy, others as a result of intralysosomal inhibition, while at least one was associated with intramitochondrial proteolysis.

1985 ◽  
Vol 232 (2) ◽  
pp. 385-393 ◽  
Author(s):  
C S Chandler ◽  
F J Ballard

Incubation of cultured cells with [3H]biotin leads to the labelling of acetyl-CoA carboxylase, pyruvate carboxylase, propionyl-CoA carboxylase and methylcrotonyl-CoA carboxylase. The biotin-containing subunits of the last two enzymes from rat cell lines are not separated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, but adequate separation is achieved with the enzymes from human cells. Since incorporated biotin is only released upon complete protein breakdown, the loss of radioactivity from gel slices coinciding with fluorograph bands was used to quantify degradation rates for each protein. In HE(39)L diploid human fibroblasts, the degradation rate constants are 0.55, 0.40, 0.31 and 0.19 day-1 for acetyl-CoA carboxylase, pyruvate carboxylase, methylcrotonyl-CoA carboxylase and propionyl-CoA carboxylase respectively. A similar series of rate constants is found for AG2804 transformed fibroblasts. The degradation rate constants are decreased by 31-67% in the presence of 50 micrograms of leupeptin/ml plus 5 mM-NH4Cl. Although the largest percentage effect was noted with the most stable enzyme, propionyl-CoA carboxylase, the absolute change in rate constant produced by the lysosomotropic inhibitors was similar for the three mitochondrial carboxylases, but greater for the cytosolic enzyme acetyl-CoA carboxylase. The heterogeneity in degradation rate constants for the mitochondrial carboxylases indicates that only part of their catabolism can occur via the autophagy-mediated unit destruction of mitochondria. Calculations showed that the autophagy-linked process had degradation rate constants of 0.084 and 0.102 day-1 respectively in HE(39)L and AG2804 cells. It accounted for two-thirds of the catabolic rate of propionyl-CoA carboxylase and a lesser proportion for the other enzymes.


2013 ◽  
Vol 304 (3) ◽  
pp. E282-E293 ◽  
Author(s):  
Charles Harris ◽  
Donald J. Roohk ◽  
Mark Fitch ◽  
Benjamin M. Boudignon ◽  
Bernard P. Halloran ◽  
...  

Glucocorticoids are extremely effective anti-inflammatory therapies, but their clinical use is limited due to severe side effects, including osteoporosis, muscle wasting, fat redistribution, and skin thinning. Here we use heavy water labeling and mass spectrometry to measure fluxes through metabolic pathways impacted by glucocorticoids. We combine these methods with measurements of body composition in corticotropin-releasing hormone (CRH)-transgenic (Tg)+ mice that have chronically elevated, endogenously produced corticosterone and a phenotype that closely mimics Cushing's disease in humans. CRH-Tg+ mice had increased adipose mass, adipose triglyceride synthesis, and greatly increased triglyceride/fatty acid cycling in subcutaneous and abdominal fat depots and increased de novo lipogenesis in the abdominal depot. In bone, CRH-Tg+ mice had decreased bone mass, absolute collagen synthesis rates, and collagen breakdown rate. In skin, CRH-Tg+ mice had decreased skin thickness and absolute collagen synthesis rates but no decrease in the collagen breakdown rate. In muscle, CRH-Tg+ mice had decreased muscle mass and absolute protein synthesis but no decrease in the protein breakdown rate. We conclude that chronic exposure to endogenous glucocorticoid excess in mice is associated with ongoing decreases in bone collagen, skin collagen, and muscle protein synthesis without compensatory reduction (coupling) of breakdown rates in skin and muscle. Both of these actions contribute to reduced protein pool sizes. We also conclude that increased cycling between triglycerides and free fatty acids occurs in both abdominal and subcutaneous fat depots in CRH-Tg+ mice. CRH-Tg mice have both increased lipolysis and increased triglyceride synthesis in adipose tissue.


2020 ◽  
Vol 18 (1) ◽  
pp. 129-137
Author(s):  
Yayuk Astuti ◽  
Rizka Andianingrum ◽  
Abdul Haris ◽  
Adi Darmawan ◽  

AbstractSynthesis of bismuth oxide synthesis through the precipitation method using H2C2O4 and Na2CO3 precipitating agents, identification of physicochemical properties and its photocatalysis activity for methyl orange degradation were conducted. The bismuth oxide synthesis was undertaken by dissolving Bi(NO3)3.5H2O in HNO3, then added precipitating agents to form precipitate. The results showed that bismuth oxide produced by H2C2O4 precipitating agent was a yellow powder containing a mixture of α-Bi2O3 (monoclinic) and β-Bi2O3 (tetragonal), porous with size of 28-85 μm. Meanwhile, the use of Na2CO3 as precipitating agent resulted in bismuth oxide consisting of α-Bi2O3 and β-Bi2O3 and Bi2O4, irregular shape without pore being 40-115 μm in size. Bismuth oxide synthesized with H2C2O4 precipitating agent showed higher photocatalytic activity compared to bismuth oxide synthesized using Na2CO3 on degrading methyl orange dye with degradation rate constants of 2.35x10-5 s-1 for H2C2O4 and 1.81x10-5 s-1 for Na2CO3.


2003 ◽  
Vol 36 (4) ◽  
pp. 428-434
Author(s):  
Takuya Kawanishi ◽  
Hiroyuki Amano ◽  
Eriko Masani ◽  
Yoshishige Hayashi ◽  
Naoto Kamata ◽  
...  

Processes ◽  
2021 ◽  
Vol 9 (11) ◽  
pp. 2025
Author(s):  
Xiaoli Dai ◽  
Jing Lv ◽  
Wenxia Wei ◽  
Shaohui Guo

High-efficiency bioremediation technology for heavy oil pollution has been a popular research topic in recent years. Laccase is very promising for the remediation of heavy oil pollution because it can not only convert bio-refractory hydrocarbons into less toxic or completely harmless compounds, but also accelerate the biodegradation efficiency of heavy oil. However, there are few reports on the use of laccase to enhance the biodegradation of heavy oil. In this study, we investigated the effect of laccase on the bacterial consortia degradation of heavy oil. The degradation efficiencies of bacterial consortia and the laccase-bacterial consortia were 60.6 ± 0.1% and 68.2 ± 0.6%, respectively, and the corresponding heavy oil degradation rate constants were 0.112 day−1 and 0.198 day−1, respectively. The addition of laccase increased the heavy oil biodegradation efficiency (p < 0.05) and biodegradation rate of the bacterial consortia. Moreover, gas chromatography–mass spectrometry analysis showed that the biodegradation efficiencies of the laccase-bacterial consortia for saturated hydrocarbons and aromatic hydrocarbons were 82.5 ± 0.7% and 76.2 ± 0.9%, respectively, which were 16.0 ± 0.3% and 13.0 ± 1.8% higher than those of the bacterial consortia, respectively. In addition, the degradation rate constants of the laccase-bacterial consortia for saturated hydrocarbons and aromatic hydrocarbons were 0.267 day−1 and 0.226 day−1, respectively, which were 1.07 and 1.15 times higher than those of the bacterial consortia, respectively. The degradation of C15 to C35 n-alkanes and 2 to 5-ring polycyclic aromatic hydrocarbons by laccase-bacterial consortia was higher than individual bacterial consortia. It is further seen that the addition of laccase significantly improved the biodegradation of long-chain n-alkanes of C22–C35 (p < 0.05). Overall, this study shows that the combination of laccase and bacterial consortia is an effective remediation technology for heavy oil pollution. Adding laccase can significantly improve the heavy oil biodegradation efficiency and biodegradation rate of the bacterial consortia.


1975 ◽  
Vol 150 (2) ◽  
pp. 235-243 ◽  
Author(s):  
D J Millward ◽  
P J Garlick ◽  
R J C Stewart ◽  
D O Nnanyelugo ◽  
J C Waterlow

Because of turnover, protein synthesis and breakdown can each be involved in the regulation of the growth of tissue protein. To investigate the regulation of skeletal-muscle-protein growth we measured rates of protein synthesis and breakdown in growing rats during development on a good diet, during development on a marginally low-protein diet and during rehabilitation on a good diet after a period of severe protein deficiency. Rates of protein synthesis were measured in vivo with a constant intravenous infusion of [14C]tyrosine. The growth rate of muscle protein was measured and the rate of breakdown calculated as breakdown rate=synthesis rate-growth rate. These measurements showed that during development on a good diet there was a fall with age in the rate of protein synthesis resulting from a fall in capacity (RNA concentration) and activity (synthesis rate per unit of RNA). There was a fall with age in the breakdown rate so that the rate was highest in the weaning rats, with a half-life of 3 days. There was a direct correlation between the fractional growth and breakdown rates. During rehabilitation on the good diet, rapid growth was also accompanied by high rates of protein breakdown. During growth on the inadequate diet protein synthesis rates were lesss than in controls, but growth occurred because of decreased rates of protein breakdown. This compression was not complete, however, since ultimate muscle size was only one-half that of controls. It is suggested that increased rates of protein breakdown are a necessary accompaniment to muscle growth and may result from the way in which myofibrils proliferate.


2009 ◽  
Vol 9 (5) ◽  
pp. 477-483 ◽  
Author(s):  
H. Tran ◽  
G. M. Evans ◽  
Y. Yan ◽  
A. V. Nguyen

Photocatalytic degradation of geosmin and 2-methylisoborneol (MIB), which are two taste and odour compounds commonly found in drinking water supply sources, was investigated using an immobilised TiO2 photoreactor. It was found that the degradation of geosmin and MIB followed similar pseudo-first-order kinetics with reaction rate constants being approximately 0.025 min−1 for typical geosmin and MIB concentrations of 250 and 500 ng/L. The normalised formal quantum efficiency was calculated to be in the range of 162–182 L/mol. Influence of additives (i.e. sodium bicarbonate and alcohols) on the degradation process was also investigated. It was found that there was a small reduction in the degradation rate constants of geosmin and MIB with increasing sodium bicarbonate concentration. At 50 mg/L sodium bicarbonate the degradation rate constants decreased by approximately 5%. Similarly, for methanol and ethanol concentrations up to 35 and 50 mg/L, respectively, these constants were found to also decrease. While addition of sodium bicarbonate and alcohols was seen to have relatively small negative effects on the photocatalytic degradation performance, the magnitude of their influence was consistent with the hypothesis that the degradation mechanism of geosmin and MIB was predominately that of attack involving HO∙ radicals.


1978 ◽  
Vol 100 (3) ◽  
pp. 386-393 ◽  
Author(s):  
E. P. Kingsbury

A testing scheme is described for short-term evaluation of lubricant-breakdown rate in starved elastrohydrodynamic (EHD) rolling contacts. It is shown that the rate is strongly influenced by the amount of lubricant available to the contacts, and by certain chemical treatments of the surfaces. Several surface conditions are evaluated for their effect on degradation rate.


Author(s):  
Philip H. Howard ◽  
Robert S. Boethling ◽  
William F. Jarvis ◽  
William M. Meylan ◽  
Edward M. Michalenko

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