A sensitive assay for ornithine amino transferase in rat brain mitochondria by ninhydrin method

Kumar, Harish; Ananda, S.; Devaraju, K. S.; Prakash, B. M.; Kumar, S. Sampath; Babu, S. V. Suresh; Ramachandraswamy, N.; Puttaraju, H. P.

doi:10.1007/s12291-009-0052-8

Cited by 5 publications

(3 citation statements)

References 23 publications

(20 reference statements)

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“…The supernatants were then used for protein estimation using the Bradford assay [ 37 ]. The activity of ornithine aminotransferase was determined using a protocol similar to that of Kumar et al [ 38 ]. The standard conditions were 35 mM l -ornithine (O2376, Sigma-Aldrich, Oakville, ON, Canada), 5 mM alpha-ketoglutarate (75890, Sigma-Aldrich, Oakville, ON, Canada), and cofactor 0.05 mM pyridoxal phosphate (82870, Sigma-Aldrich, Oakville, ON, Canada) in 50 mM Tris-HCl (pH 8.0) of a total volume of 1 mL.…”

Section: Methodsmentioning

confidence: 99%

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Breast Cancer-Derived Microvesicles Are the Source of Functional Metabolic Enzymes as Potential Targets for Cancer Therapy

et al. 2021

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Membrane-derived extracellular vesicles, referred to as microvesicles (MVs), have been proposed to participate in several cancer diseases. In this study, MV fractions were isolated by differential ultracentrifugation from a metastatic breast cancer (BC) cell line MDA-MB-231 and a non-cancerous breast cell line MCF10A, then analyzed by nano-liquid chromatography coupled to tandem mass spectrometry. A total of 1519 MV proteins were identified from both cell lines. The data obtained were compared to previously analyzed proteins from small extracellular vesicles (sEVs), revealing 1272 proteins present in both MVs and sEVs derived from the MDA-MB-231 cell line. Among the 89 proteins unique to MDA-MB-231 MVs, three enzymes: ornithine aminotransferase (OAT), transaldolase (TALDO1) and bleomycin hydrolase (BLMH) were previously proposed as cancer therapy targets. These proteins were enzymatically validated in cells, sEVs, and MVs derived from both cell lines. The specific activity of OAT and TALDO1 was significantly higher in MDA-MB-231-derived MVs than in MCF10A MVs. BLMH was highly expressed in MDA-MB-231-derived MVs, compared to MCF10A MVs. This study shows that MVs carry functional metabolic enzymes and provides a framework for future studies of their biological role in BC and potential in therapeutic applications.

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Section: Methodsmentioning

confidence: 99%

“…The reaction mixture was incubated at 37 °C for 30 min. The reaction was stopped by adding 0.25 mL of 3.6 N perchloric acid (311421, Sigma-Aldrich, Oakville, ON, Canada) and 0.25 mL of 2% ninhydrin (151173, Sigma-Aldrich, Oakville, ON, Canada) in water and proceeded as described previously [ 38 ]. Controls were stopped at time 0.…”

Section: Methodsmentioning

confidence: 99%

Breast Cancer-Derived Microvesicles Are the Source of Functional Metabolic Enzymes as Potential Targets for Cancer Therapy

et al. 2021

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“…[39][40][41][42][43][44][45] Ninhydrin is extensively used to detect compounds of pharmaceutical importance [46][47][48][49][50][51][52][53][54][55][56][57] and in kinetic studies. [58][59] Scheme 1.…”

Section: -10mentioning

confidence: 99%

Ninhydrin in synthesis of heterocyclic compounds

Ziarani¹,

Lashgari²,

Azimian³

et al. 2015

Arkivoc

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Ninhydrin has been utilized in many heterocyclic preparations and considered as an important building block in organic synthesis. There is a wide range of reactions that include ninhydrin in the synthesis of heterocyclic compounds. This review highlights the advances in the use of ninhydrin as starting material in the synthesis of various organic compounds and drugs in a fully comprehensive way, from its first isolation in 1910 to the end of 2013. There is also a diversity of multi-component reactions of ninhydrin and we highlight the recent reports in this review.

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Underestimated contribution of skeletal muscle in ornithine metabolism during mouse postnatal development

2013

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Ornithine aminotransferase (L-ornithine 2-oxoacid aminotransferase, OAT) is widely expressed in organs, but studies in mice have focused primarily on the intestine, kidney and liver because of the high OAT-specific activity in these tissues. This study aimed to investigate OAT activity in adult mouse tissues to assess the potential contribution to ornithine metabolism and to determine OAT control during postnatal development. OAT activity was widely distributed in mouse tissues. Sexual dimorphism was observed for most tissues in adults, with greater activity in females than in males. The contribution of skeletal muscles to total OAT activity (34% in males and 27% in females) was the greatest (50%) of the investigated tissues in pre-weaned mice and was similar to that of the liver in adults. OAT activity was found to be regulated in a tissue-specific manner during postnatal development in parallel with large changes in the plasma testosterone and corticosterone levels. After weaning, OAT activity markedly increased in the liver but dropped in the skeletal muscle and adipose tissue. Anticipating weaning for 3 days led to an earlier reduction of OAT activity in skeletal muscles. Orchidectomy in adults decreased OAT activity in the liver but increased it in skeletal muscle and adipose tissue. We concluded that the contribution of skeletal muscle to mouse ornithine metabolism may have been underestimated. The regulation of OAT in skeletal muscles differs from that in the liver. The present findings suggest important and tissue-specific metabolic roles for OAT during postnatal development in mice.

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A sensitive assay for ornithine amino transferase in rat brain mitochondria by ninhydrin method

Abstract: ABSTRACT

Cited by 5 publications

References 23 publications

Breast Cancer-Derived Microvesicles Are the Source of Functional Metabolic Enzymes as Potential Targets for Cancer Therapy

Breast Cancer-Derived Microvesicles Are the Source of Functional Metabolic Enzymes as Potential Targets for Cancer Therapy

Ninhydrin in synthesis of heterocyclic compounds

Underestimated contribution of skeletal muscle in ornithine metabolism during mouse postnatal development

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