Rat liver pyruvate carboxylase. Purification, detection and quantification of apo and holo forms by immuno-blotting and by an enzyme-linked immunosorbent assay

Ahmad, Fazal; Ahmad, Patricia M.; Méndez, Alma Pérez

doi:10.1042/bj2360527

Cited by 9 publications

(5 citation statements)

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“…Genetic variations among the circulating norovirus strains along with inter-genotype recombinant viruses have been reported in India as well [ 13 , 14 ]. GII.P16-GII.13 was reported previously by Nataraju et al [ 25 ] in eastern India, and this genotype has also been found in Spain, Germany, and Greece [ 55 , 58 , 59 ]. During this study period too, recombinant viruses were identified in a few patients admitted with AGE.…”

Section: Discussionsupporting

confidence: 68%

Genetic characterization and evolutionary analysis of norovirus genotypes circulating among children in eastern India during 2018-2019

Mitra

et al. 2021

Arch Virol

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Noroviruses are significant etiological agents of acute gastroenteritis (AGE) across all age groups, especially in children under 5 years of age. Although the prevalence of norovirus infection is known to have increased in various countries, in India there are few reports pertaining to the norovirus disease burden. We investigated the epidemiology and molecular characteristics of noroviruses in children seeking health care at two hospitals in Kolkata, Eastern India. Faecal specimens were collected between January 2018 and December 2019 from 2812 children under 5 years of age with acute gastroenteritis. Noroviruses were detected in 6.04% (170/2812) of the samples, and 12.9% (22/170) of these were cases of coinfection with rotavirus. Among children (≤5 years), a higher infection rate (8.2%, n = 94/1152) was observed in the 6 to 12 month age group. GII.4 Sydney 2012 was the dominant norovirus capsid genotype (n = 75/90, 83.3%), followed by GII.3 (n = 10/90, 11.1%). Other capsid types GII.13 (n = 4/90, 4.4%) and GII.17 (n = 1/90; 1.1%) were also detected at low frequency. Phylogenetic analysis showed that the GII.P16 polymerase of strains in this region clustered with those of the phylogenetically distinct monophyletic clade of GII.P16 strains, whose members have been circulating worldwide since 2014. Inter-genotypic norovirus recombinants such as GII.P16-GII.3 (n = 10) and GII.P16-GII.13 (n = 4) were also observed among the circulating strains. In comparison to previous studies from eastern India, the present study shows a higher detection rate of norovirus infection in the paediatric population suffering from acute gastroenteritis. Continuous surveillance is required for predicting the emergence of novel genotypes and recombinant strains and for future vaccine development. Supplementary Information The online version contains supplementary material available at 10.1007/s00705-021-05197-6.

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Section: Discussionsupporting

confidence: 68%

Genetic characterization and evolutionary analysis of norovirus genotypes circulating among children in eastern India during 2018-2019

Mitra

et al. 2021

Arch Virol

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“…We considered the possibility that PC of cAMP-treated adipocytes may be inactive because of the subtraction or lack of incorporation of its biotin prosthetic group. In previous studies we showed that apo-and holo-PC and holo-PC-avidin complex react similarly with anti-PC antibodies (Ahmad et al, 1986). This procedure provided a convenient method for the simultaneous detection and quantification of apo-and/or holo-forms of PC in cell homogenates.…”

Section: Figurementioning

confidence: 79%

“…PC activity was measured at 37°C by a radioisotopic C02fixation assay in the presence of Triton X-100 (Atkin et al, 1979) as described (Ahmad et al, 1986); 1 munit of enzyme activity represents the synthesis of 1 nmol of oxaloacetate/min and the specific activity is defined as munits/mg of protein.…”

Section: Enzyme Assaysmentioning

confidence: 99%

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Regulation of pyruvate carboxylase in 3T3-L1 cells

Zhang

Xia

Ahmad

1995

Biochemical Journal

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When 3T3-L1 fibroblasts differentiate to adipocytes, the specific activity of pyruvate carboxylase (PC) increases about 25-fold in parallel with its intracellular protein concentration. The increase in PC protein concentration is accompanied by a 9-10-fold increase in the relative abundance of 4.2 kb PC mRNA measured by Northern-blot analysis using a cDNA probe encoding a segment of the PC gene of 3T3-L1 adipocytes. The effects of cyclic AMP (cAMP) alone and together with insulin on levels of cellular protein, PC activity, PC protein and on the relative abundance of PC mRNA were examined in mature 3T3-L1 adipocytes. Adipocytes exposed to cAMP for 24 h exhibited a 25% decrease in cellular protein and marked decreases in enzyme activity (88%) and PC mRNA abundance (98%) compared with untreated adipocyte controls. After 48 h of exposure to cAMP, PC activity and PC mRNA diminished to levels approaching their detection limits. When exposed to medium containing cAMP plus insulin, adipocyte enzyme activity and PC mRNA declined more slowly during the first 24 h exposure (about 20% decrease) but after 48 h fell to values comparable with those of adipocytes exposed to cAMP alone. Despite these decreases in enzyme activity, the PC protein content of adipocytes treated with cAMP alone or cAMP plus insulin are nearly identical with that of control adipocytes. The inactivation of PC in cAMP-treated adipocytes does not involve loss of the prosthetic group from the holoenzyme. Cross-linking experiments suggest that the spatial arrangement of protomers in inactive PC may differ from that in the active tetrameric enzyme. Data presented suggest that, in addition to inducing inactivation, cAMP may also regulate adipocyte PC by decreasing transcription of the PC gene and/or enhancing the rate of degradation of PC mRNA.

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“…Specific activity is expressed in units/mg protein. The amount of PC was assayed by a competitive ELISA according to Ahmad et al (1986). Different standard calibration curves with purified rat-liver or rat-kidney cortex PC were used.…”

Section: Animals and Treatmentsmentioning

confidence: 99%

Effects of Starvation, Diabetes and Carbon Tetrachloride Intoxication on Rat Kidney Cortex and Liver Pyruvate Carboxylase Levels

Salto¹,

Sola²,

Oliver³

et al. 1996

Archives of Physiology and Biochemistry

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Rat liver pyruvate carboxylase. Purification, detection and quantification of apo and holo forms by immuno-blotting and by an enzyme-linked immunosorbent assay

Cited by 9 publications

References 35 publications

Genetic characterization and evolutionary analysis of norovirus genotypes circulating among children in eastern India during 2018-2019

Genetic characterization and evolutionary analysis of norovirus genotypes circulating among children in eastern India during 2018-2019

Regulation of pyruvate carboxylase in 3T3-L1 cells

Effects of Starvation, Diabetes and Carbon Tetrachloride Intoxication on Rat Kidney Cortex and Liver Pyruvate Carboxylase Levels

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