A method usingL/CMS for determination of absolute configuration of constituent amino acids in peptide --- advanced Marfey's method ---

Harada, Kenichi; Fujii, Kiyonaga; Mayumi, Tsuyoshi; Hibino, Yasuko; Suzuki, Makoto; Ikai, Yoshitomo; Oka, Hisao

doi:10.1016/0040-4039(95)00078-q

Cited by 78 publications

(103 citation statements)

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“…In order to determine the stereochemistry of the Ahp moiety, 1 was subjected to oxidation using PCC/ CH 2 Cl 2 followed by acid hydrolysis and Marfey's analysis as outlined above. The exclusive presence of L-glutamic acid in the hydrolysate showed that the Ahp moiety also possessed the L-configuration.Using the advanced Marfey's method with HPLC-MS analysis 9 we were able to demonstrate that the novel amino acid N,O-dimethylbromotyrosine also possessed an L-configuration. Direct Marfey's analysis using LC-MS identified an HPLC peak of m/z 568/570 with a retention time of 64.8 min.…”

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confidence: 99%

Symplocamide A, a Potent Cytotoxin and Chymotrypsin Inhibitor from the Marine Cyanobacterium Symploca sp.

et al. 2007

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Investigation of a Symploca sp. from Papua New Guinea has led to the isolation of symplocamide A (1), a potent cancer cell cytotoxin, which also inhibits serine proteases with a 200-fold greater inhibition of chymotrypsin over trypsin. The complete stereostructure of symplocamide A was determined by detailed NMR and MS analysis as well as chiral HPLC analysis of the component amino acid residues. The presence of several unusual structural features in symplocamide A provides new insights into the pharmacophore model for protease selectivity in this drug class and may underlie the potent cytotoxicity of this compound to H-460 lung cancer cells (IC 50 = 40 nM) as well as neuro-2a neuroblastoma cells (IC 50 = 29 nM).Marine cyanobacteria are a continuing source of many novel natural product structures, some of which possess highly potent biological properties. 1 Most of these metabolites are partially composed of amino acids, and these are often integrated with polyketide sections to make a wide range of nitrogen-rich lipids. These lipopeptide backbones are often modified by oxidation, methylation, or various halogenations, including those catalyzed by the recently described radical halogenases. 2,3 There is a growing body of literature that supports the hypothesis that the presence of halogen atoms in natural products enhances their biological potency. 4,5 Hence, we have been interested in isolating new types of halogenated lipopeptides from marine cyanobacteria with the goals of further defining the extent of their structural diversity and biological properties.Proteases are implicated in the pathogenesis of many human diseases, including cancer and Alzheimer's, and thus the therapeutic modulation of proteolytic activity offers an attractive potential treatment modality. Indeed, it has been estimated that of the approximately 400 human *To whom correspondence should be addressed. Tel: (858) 6 In addition, proteases from infectious pathogens such as HIV, HCV, and dengue virus are crucial drug targets for the improvement of human health. However, with this multitude of different proteases and many potential therapeutic applications, discovery of agents with selectivity for specific proteases is crucial to the development of truly useful pharmaceuticals in this class.While freshwater cyanobacteria are a rich source of structurally diverse protease inhibitors, 7 their marine relatives represent an under-explored resource of this enzyme-inhibitor class. Hence, we have initiated a program to survey marine cyanobacterial extracts, fractions, and newly isolated pure compounds for interesting profiles of protease inhibition. This line of research has been productive and led to the identification of the depsipeptide symplocamide A (1), which shows strong serine protease inhibitory activity. In addition, symplocamide A shows a high level of cytotoxicity to cancer cells in vitro (H-460 lung cancer cells, IC 50 = 40 nM; neuro-2a neuroblastoma cells, IC 50 = 29 nM). These biological properties combined with its unusual s...

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confidence: 99%

Symplocamide A, a Potent Cytotoxin and Chymotrypsin Inhibitor from the Marine Cyanobacterium Symploca sp.

et al. 2007

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“…General 1 H and 13 C NMR spectra were recorded at 298 K on an AVANCE 500 (Bruker Biospin) spectrometer operating at 500 MHz and 125 MHz, respectively. The samples for NMR characterization were dissolved in chloroform-d or methanol-d 4 .…”

Section: Methodsmentioning

confidence: 99%

“…High-resolution mass spectra were measured with a JMS700 (JEOL) or LCT. Mass spectra for advanced Marfey's method [13] were recorded with an LCQ ion-trap mass spectrometer equipped with an electrospray ionization (ESI) probe (Thermo Electron Corporation) coupled on-line to an HP1100 system (Agilent Technology).…”

Section: Methodsmentioning

confidence: 99%

“…2) Pleofungin B The 13 C and 1 H NMR data (CDCl 3 ) of pleofungin B are shown in Table 3. The molecular weight of pleofungin B was determined to be 1,136 by FAB-MS.…”

Section: Physico-chemical Propertiesmentioning

confidence: 99%

“…1. 3) Pleofungin C The 13 C and 1 H NMR data (CDCl 3 ) of pleofungin C are shown in Table 4. The molecular weight of pleofungin C was determined to be 1,080 and the molecular formula was established as C 54 H 96 N 8 O 14 from the result of HRTOF-MS. Amino acid analysis of its acid hydrolysate (6 M HCl, 105°C, 12 hours) showed the existence of Ala, Thr and Val.…”

Section: Physico-chemical Propertiesmentioning

confidence: 99%

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Pleofungins, Novel Inositol Phosphorylceramide Synthase Inhibitors, from Phoma sp. SANK 13899

Aoyagi

Yano²,

Kozuma³

et al. 2007

J Antibiot

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Pleofungins (formerly called F-15078) A, B, C and D, novel depsipeptide antifungal antibiotics, were found in a mycelium extract of the producing fungus, Phoma sp. SANK 13899. The structures of pleofungins A, B, C and D were elucidated mainly by various NMR studies. The absolute configurations of the amino acids and N-methyl amino acids of pleofungin A constituents in the hydrolysate were determined by the application of advanced Marfey's method in combination with gas chromatography/mass spectrometry analysis of their silylation products with N-methyl-N-(tert-butylsilyl)-trifluoroacetamide. Two a -hydroxy acid constituents, ahydroxyisocaproic acid and a-hydroxyisovaleric acid, were isolated from the hydrolysate and their stereochemistries were determined by their specific rotations.

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HPLC determination of acidic d‐amino acids and their N‐methyl derivatives in biological tissues

Tsesarskaia

Galindo

Szókán

et al. 2009

Biomedical Chromatography

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d-aspartate (d-Asp) and N-methyl-d-aspartate (NMDA) occur in the neuroendocrine systems of vertebrates and invertebrates where they play a role in hormone release and synthesis, neurotransmission, and memory and learning. N-methyl-d-glutamate (NMDG) has also been detected in marine bivalves. Several methods have been used to detect these amino acids, but they require pretreatment of tissue samples with o-phthaldialdehyde (OPA) to remove primary amino acids which interfere with the detection of NMDA and NMDG. We report here a one step derivatization procedure with the chiral reagent N-α-(5-fluoro-2,4-dinitrophenyl)-(d or l)-valine amide, FDNP-Val-NH2, a close analog of Marfey’s reagent but with better resolution and higher molar absorptivity. The diastereomers formed are separated by HPLC on an ODS-Hypersil column eluted with TFA/water – TFA/MeCN. UV absorption at 340 nm permits detection levels as low as 5–10 picomoles. D-Asp, NMDA and NMDG peaks are not obscured by other primary or secondary amino acids; hence pretreatment of tissues with OPA is not required. This method is highly reliable and fast (less than 40 minutes HPLC run). Using this method, we have detected D-Asp, NMDA and NMDG in several biological tissues (octopus brain, optical lobe, and bucchal mass; foot and mantle of the mollusk Scapharca broughtonii), confirming the results of other researchers.

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A method usingL/CMS for determination of absolute configuration of constituent amino acids in peptide --- advanced Marfey's method ---

Cited by 78 publications

References 6 publications

Symplocamide A, a Potent Cytotoxin and Chymotrypsin Inhibitor from the Marine Cyanobacterium Symploca sp.

Symplocamide A, a Potent Cytotoxin and Chymotrypsin Inhibitor from the Marine Cyanobacterium Symploca sp.

Pleofungins, Novel Inositol Phosphorylceramide Synthase Inhibitors, from Phoma sp. SANK 13899

HPLC determination of acidic d‐amino acids and their N‐methyl derivatives in biological tissues

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