Quantitating adenylate nucleotides in diverse organisms

Napolitano, Michael J.; Shain, Daniel H.

doi:10.1016/j.jbbm.2005.03.001

Cited by 30 publications

(27 citation statements)

References 17 publications

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“…Nucleotides were extracted according to Napolitano and Shain (2005). Fifty microliters of chloroplast solution were mixed with 50 mL of 7% (v/v) perchloric acid and incubated for 10 min at 4°C.…”

Section: Extraction and Detection Of Nucleotidesmentioning

confidence: 99%

Chloroplastic Phosphoadenosine Phosphosulfate Metabolism Regulates Basal Levels of the Prohormone Jasmonic Acid in Arabidopsis Leaves

et al. 2010

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Levels of the enzymes that produce wound response mediators have to be controlled tightly in unwounded tissues. The Arabidopsis (Arabidopsis thaliana) fatty acid oxygenation up-regulated8 ( fou8) mutant catalyzes high rates of a-linolenic acid oxygenation and has higher than wild-type levels of the a-linolenic acid-derived wound response mediator jasmonic acid (JA) in undamaged leaves. fou8 produces a null allele in the gene SAL1 (also known as FIERY1 or FRY1). Overexpression of the wild-type gene product had the opposite effect of the null allele, suggesting a regulatory role of SAL1 acting in JA synthesis. The biochemical phenotypes in fou8 were complemented when the yeast (Saccharomyces cerevisiae) sulfur metabolism 3#(2#), 5#-bisphosphate nucleotidase MET22 was targeted to chloroplasts in fou8. The data are consistent with a role of SAL1 in the chloroplast-localized dephosphorylation of 3#-phospho-5#-adenosine phosphosulfate to 5#-adenosine phosphosulfate or in a closely related reaction (e.g. 3#,5#-bisphosphate dephosphorylation). Furthermore, the fou8 phenotype was genetically suppressed in a triple mutant ( fou8 apk1 apk2) affecting chloroplastic 3#-phospho-5#-adenosine phosphosulfate synthesis. These results show that a nucleotide component of the sulfur futile cycle regulates early steps of JA production and basal JA levels.

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Section: Extraction and Detection Of Nucleotidesmentioning

confidence: 99%

Chloroplastic Phosphoadenosine Phosphosulfate Metabolism Regulates Basal Levels of the Prohormone Jasmonic Acid in Arabidopsis Leaves

et al. 2010

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“…Adenosine triphosphate (ATP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP) were measured in neutralized perchloric acid extracts of frozen ground samples using a luciferin-luciferase method (Napolitano and Shain, 2005) and a luminescence ATP detection kit (PerkinElmer TM , Zaventem, Belgium). The extraction was performed twice for each position, to be used as technological replicates.…”

Section: Primary Metabolites and Adenine Nucleotide Quantificationmentioning

confidence: 99%

Metabolic acclimation to hypoxia revealed by metabolite gradients in melon fruit

Biais

Beauvoit

Allwood

et al. 2010

Journal of Plant Physiology

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“…ATP was extracted as described elsewhere. 7 Briefly, cells were spun at 2000 UPM before lysed with proteinase K in HEPES buffer, pH 7.4. After 15 min of incubation at 501C, proteinase K was heat-inactivated at 801C and cell lysates were frozen in liquid nitrogen.…”

Section: Investigation Of the Mitochondrial Membrane Potentialmentioning

confidence: 99%

Mechanisms of apoptosis-induction by rottlerin: therapeutic implications for B-CLL

et al. 2006

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Constitutively activated signaling pathways contribute to the apoptosis-defect of B-CLL cells. Protein kinase C-d is a permanently activated kinase and a putative downstream target of phosphatidylinositol-3 kinase in B-CLL. Blockade of protein kinase C-d (PKC-d) by the highly specific inhibitor rottlerin induces apoptosis in chronic lymphocytic leukaemia (CLL) cells. By co-culturing bone marrow stromal and CLL cells, we determined that the proapoptotic effect of rottlerin is not abolished in the presence of survival factors, indicating that a targeted therapy against PKC-d might be a powerful approach for the treatment of CLL patients. The downstream events following rottlerin treatment engage mitochondrial and nonmitochondrial pathways and ultimately activate caspases that execute the apoptotic cell death. Herein we report that the inhibition of PKC-d decreases the expression of the important antiapoptotic proteins Mcl-1 and XIAP accompanied by a loss of the mitochondrial membrane potential Dw. In addition, we discovered that ZAP-70-expressing cells are significantly more susceptible to rottlerin-induced cell death than ZAP-70 negative cells. We finally observed that rottlerin can augment cell toxicity induced by standard chemotherapeutic drugs. Conclusively, PKC-d is a promising new target in the combat against CLL.

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Quantitating adenylate nucleotides in diverse organisms

Cited by 30 publications

References 17 publications

Chloroplastic Phosphoadenosine Phosphosulfate Metabolism Regulates Basal Levels of the Prohormone Jasmonic Acid in Arabidopsis Leaves

Chloroplastic Phosphoadenosine Phosphosulfate Metabolism Regulates Basal Levels of the Prohormone Jasmonic Acid in Arabidopsis Leaves

Metabolic acclimation to hypoxia revealed by metabolite gradients in melon fruit

Mechanisms of apoptosis-induction by rottlerin: therapeutic implications for B-CLL

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