Colorimetric Assay Methods for Free and Phosphorylated Glyceric Acids

Bartlett, Grant R.

doi:10.1016/s0021-9258(18)70227-5

Cited by 361 publications

(41 citation statements)

References 2 publications

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“…Glyceric acid was determined either by colorimetric estimation of formaldehyde liberated from C-3 by periodate (Frisell, Meech & Mackenzie, 1954) or by the colour reaction with chromotropic acid in nearly conc. H2SO4 (Bartlett, 1959). In the latter assay, for a given amount of glyceric acid, maximum colour production occurred when the chromotropic acid concentration was 0-025%, and the method was modified accordingly.…”

Section: Methodsmentioning

confidence: 99%

THE OXIDATION OF d- AND l-GLYCERATE BY RAT LIVER

Dawkins¹,

Dickens²

1965

Biochemical Journal

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1. The interconversion of hydroxypyruvate and l-glycerate in the presence of NAD and rat-liver l-lactate dehydrogenase has been demonstrated. Michaelis constants for these substrates together with an equilibrium constant have been determined and compared with those for pyruvate and l-lactate. 2. The presence of d-glycerate dehydrogenase in rat liver has been confirmed and the enzyme has been purified 16-20-fold from the supernatant fraction of a homogenate, when it is free of l-lactate dehydrogenase, with a 23-29% recovery. The enzyme catalyses the interconversion of hydroxypyruvate and d-glycerate in the presence of either NAD or NADP with almost equal efficiency. d-Glycerate dehydrogenase also catalyses the reduction of glyoxylate, but is distinct from l-lactate dehydrogenase in that it fails to act on pyruvate, d-lactate or l-lactate. The enzyme is strongly dependent on free thiol groups, as shown by inhibition with p-chloromercuribenzoate, and in the presence of sodium chloride the reduction of hydroxypyruvate is activated. Michaelis constants for these substrates of d-glycerate dehydrogenase and an equilibrium constant for the NAD-catalysed reaction have been calculated. 3. An explanation for the lowered V(max.) with d-glycerate as compared with dl-glycerate for the rabbit-kidney d-alpha-hydroxy acid dehydrogenase has been proposed.

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Section: Methodsmentioning

confidence: 99%

THE OXIDATION OF d- AND l-GLYCERATE BY RAT LIVER

Dawkins¹,

Dickens²

1965

Biochemical Journal

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“…The ANTS/DPX containing vesicles were separated from free ANTS/DPX by exclusion chromatography using a column filled with Sephadex™ G-75 (Amesham Biosciences) fine gel swollen in an isosmotic buffer (10 mM HEPES, 140 mM NaCl, pH 7.4). The void volume fractions were collected, and the phospholipid concentration was determined by phosphate analysis [76,77]. The liposomes containing ANTS/DPX were stored in the dark at room temperature.…”

Section: Cell Lines and Cell Culturementioning

confidence: 99%

Effect of L- to D-Amino Acid Substitution on Stability and Activity of Antitumor Peptide RDP215 against Human Melanoma and Glioblastoma

Maxian

Gerlitz

Riedl

et al. 2021

IJMS

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The study investigates the antitumor effect of two cationic peptides, R-DIM-P-LF11-215 (RDP215) and the D-amino acid variant 9D-R-DIM-P-LF11-215 (9D-RDP215), targeting the negatively charged lipid phosphatidylserine (PS) exposed by cancer cells, such as of melanoma and glioblastoma. Model studies mimicking cancer and non-cancer membranes revealed the specificity for the cancer-mimic PS by both peptides with a slightly stronger impact by the D-peptide. Accordingly, membrane effects studied by DSC, leakage and quenching experiments were solely induced by the peptides when the cancer mimic PS was present. Circular dichroism revealed a sole increase in β-sheet conformation in the presence of the cancer mimic for both peptides; only 9D-RDP215 showed increased structure already in the buffer. Ex vitro stability studies by SDS-PAGE as well as in vitro with melanoma A375 revealed a stabilizing effect of D-amino acids in the presence of serum, which was also confirmed in 2D and 3D in vitro experiments on glioblastoma LN-229. 9D-RDP215 was additionally able to pass a BBB model, whereupon it induced significant levels of cell death in LN-229 spheroids. Summarized, the study encourages the introduction of D-amino acids in the design of antitumor peptides for the improvement of their stable antitumor activity.

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“…The unencapsulated calcein was removed from the SUV dispersion by the Sephadex G65 mini-column separation technique (Fu and Singh, 1999). The actual phospholipid content of each preparation was determined by phosphorus assay (Bartlett, 1958) and the concentration of liposomes was adjusted for each type of experiment to 5 μM in 10 mM TRIS-HCl buffer at pH 7.4.…”

Section: Calcein Leakagementioning

confidence: 99%

Is Exogenous Fatty Acid Hydroperoxide Perception as Elicitor Related to Modulation of Plant Plasma Membrane Structure ?

Deboever

Aubel²,

Rondelli

et al. 2021

Preprint

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Oxylipins are lipid-derived molecules that are ubiquitous in eukaryotes and whose functions in plant physiology have been widely reported. They appear to play a major role in plant immunity by orchestrating reactive oxygen species (ROS) and hormone-dependent signalling pathways. The present work focuses on the specific case of fatty acid hydroperoxides (HPOs). Although some studies report their potential use as exogenous biocontrol agents for plant protection, evaluation of their efficiency in planta is lacking and no information is available about their mechanism of action. In this work, the potential of 13(S)-hydroperoxyoctadeca-(9Z,11E)-dienoic acid (13-HPOD) and 13(S)-hydroperoxy-(9Z,11E,15Z)-octadecatrienoic acid (13-HPOT), as plant defence elicitors and the underlying mechanism of action are investigated. Arabidopsis thaliana leaf resistance to Botrytis cinerea was observed after root application with HPOs. They also activate early immunity-related defence responses, like ROS. As previous studies have demonstrated their ability to interact with plant plasma membranes (PPM), we have further investigated the effects of HPOs on biomimetic PPM structure using complementary biophysics tools. Results show that HPO insertion into PPM impacts its global structure without solubilizing it. Relationship between biological assays and biophysical analysis suggests that lipid amphiphilic elicitors that directly act on membrane lipids might trigger early plant defence events

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Colorimetric Assay Methods for Free and Phosphorylated Glyceric Acids

Cited by 361 publications

References 2 publications

THE OXIDATION OF d- AND l-GLYCERATE BY RAT LIVER

THE OXIDATION OF d- AND l-GLYCERATE BY RAT LIVER

Effect of L- to D-Amino Acid Substitution on Stability and Activity of Antitumor Peptide RDP215 against Human Melanoma and Glioblastoma

Is Exogenous Fatty Acid Hydroperoxide Perception as Elicitor Related to Modulation of Plant Plasma Membrane Structure ?

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