Improved method for direct high-performance liquid chromatography assay of angiotensin-converting enzyme-catalyzed reactions

Wu, Jianping; Aluko, Rotimi E.; Muir, Alister D.

doi:10.1016/s0021-9673(02)00052-3

Cited by 149 publications

(103 citation statements)

References 19 publications

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“…The method was according to Wu, Aluko and Muir (2002) with some modifications. The total reaction volume was 225 µL, made up of 50 µL of 5 mM HHL, 160 µL of 0.025 U/mL of ACE and 20 µL of sample at different concentration (all prepared with 100 mM potassiumphosphate buffer, containing 300 mM NaCl, pH 8.3).…”

Section: Ace Inhibitionmentioning

confidence: 99%

“…In connection with this, Wu et al (2002) have reported that the spectrophotometric assay described by Cushman and Cheung (1971), widely used for testing ACE inhibition of gelatin hydrolysates, tends to overestimate the amount of HA produced during ACE-catalyzed reactions. Having this in mind, IC 50 values of the most potent inhibitor (peptide I), were between 26-and 1.5-fold higher than those reported for ACE inhibitory peptides derived from different food proteins, tested in similar conditions (Centeno et al, 2006;He, Chen, Sun, Zhang & Zhou, 2006;Tsai, Lin, Chen, & Pan, 2006;Tauzin, Miclo, & Gaillard, 2002).…”

Section: Ace Inhibitory Activity Of Synthetically Derived Peptidesmentioning

confidence: 99%

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Contribution of Leu and Hyp residues to antioxidant and ACE-inhibitory activities of peptide sequences isolated from squid gelatin hydrolysate

et al. 2011

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** This centre has implemented and maintains a Quality Management System which fulfils the requirements of the ISO standard 9001:2000 AbstractSquid gelatin obtained from inner and outer tunics was hydrolyzed with Alcalase to isolate antioxidant peptide sequences. The ACE-inhibitory activity of the isolated peptides was also evaluated. After fractionation by ultrafiltration and size-exclusion chromatography into four fractions, the antioxidant activity of the peptide fractions was determined by radical scavenging ability and ferric reducing power. Fraction FIII showed the highest antioxidant activity, although slight differences could be expected in the antioxidant activity of the different fractions based on the amino acid composition.FIII was subjected to liquid chromatography and tandem mass spectrometry (LC-MS/MS) and two major compounds were identified: the compound with m/z 952.42, which could be mostly comprised by the carbohydrate fucose, and the peptide with m/z 1410.63. Three possible sequences were proposed for this peptide, and the contribution of Leu or Hyp residues to the antioxidant and ACE-inhibitory activities of the resulting sequence was evaluated. The presence of Leu residues in the peptide sequence in *Manuscript Click here to view linked References 2 replacement of Hyp seems to play an important role in the antioxidant and ACEinhibitory activity.

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Section: Ace Inhibitionmentioning

confidence: 99%

Section: Ace Inhibitory Activity Of Synthetically Derived Peptidesmentioning

confidence: 99%

Contribution of Leu and Hyp residues to antioxidant and ACE-inhibitory activities of peptide sequences isolated from squid gelatin hydrolysate

et al. 2011

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“…O preparo das amostras para a avaliação da atividade inibitória dos hidrolisados enzimáticos de WPC sobre a ECA foi realizada de acordo com Wu et al (2002). Assim, um volume de 12,5 µL de solução de HHL (2,17 mmol.L -1 ) foi misturado com 50 µL das soluções de hidrolisados (10 mg.mL -1 ), ambas preparadas em tampão borato a 100 mmol.L -1 , contendo solução de NaCl a 300 mmol.L -1 , em pH 8,3, e a mistura foi incubada a 37 ºC por 10 minutos.…”

Section: Avaliação In Vitro Da Atividade Inibitó-ria Dos Hidrolisadosunclassified

OBTENÇÃO DE HIDROLISADOS DE CONCENTRADO PROTEICO DE SORO DE LEITE COM ATIVIDADE INIBITÓRIA DA ENZIMA CONVERSORA DE ANGIOTENSINA: AÇÃO DAS PROTEASES DO Bacillus licheniformis E DO Aspergillus oryzae

SILVESTRE¹,

Morais²,

SILVA³

et al. 2011

Publ.Exatas

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“…The ability of EcDCP to cleave dipeptide from the Cterminus of HHL was determined as described previously (Wu et al 2002) with some modifications. The reaction mixture in a total volume of 250 µL contained 30 mM universal buffer (pH 7.0), 2.5 mM HHL, 200 mM NaCl, and 20 µL of the enzyme solution at a suitable dilution.…”

Section: Enzyme Assaymentioning

confidence: 99%

Probing the catalytically essential residues of a recombinant dipeptidyl carboxypeptidase from Escherichia coli

Chen

Yen

et al. 2010

Biologia

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In studying the structure and function of Escherichia coli dipeptidyl carboxypeptidase (EcDCP), we have employed in vitro mutagenesis and subsequent protein expression to genetically dissect the enzyme in order to gain insight into the catalytic mechanism. Comparison of the amino acid sequence of EcDCP with other homologues indicates that the active site of the enzyme exhibits an HEXXH motif, a common feature of zinc metalloenzymes. The third metal binding ligand, presumed to coordinate directly to the active-site zinc ion in concert with His470 and His474 has been proposed as Glu499. Alterations to these residues completely abolished the catalytic activity against N-benzoyl-L-glycyl-L-histidyl-L-leucine. A significant loss of the enzymatic activity was also observed in F472V and F500V mutant enzymes. Intrinsic tryptophan fluorescence revealed the significant alterations of the microenvironment of aromatic amino acid residues in all mutant enzymes, whereas circular dichroism spectra were nearly identical for the tested proteins. Computer modeling suggests that residues His470, Glu471, His474, Glu499, and Phe500 are essential for EcDCP in maintaining the stable active-site environment. Taken together, these studies contribute to a more comprehensive understanding of the catalytic mechanism of the enzyme.

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Improved method for direct high-performance liquid chromatography assay of angiotensin-converting enzyme-catalyzed reactions

Cited by 149 publications

References 19 publications

Contribution of Leu and Hyp residues to antioxidant and ACE-inhibitory activities of peptide sequences isolated from squid gelatin hydrolysate

Contribution of Leu and Hyp residues to antioxidant and ACE-inhibitory activities of peptide sequences isolated from squid gelatin hydrolysate

OBTENÇÃO DE HIDROLISADOS DE CONCENTRADO PROTEICO DE SORO DE LEITE COM ATIVIDADE INIBITÓRIA DA ENZIMA CONVERSORA DE ANGIOTENSINA: AÇÃO DAS PROTEASES DO Bacillus licheniformis E DO Aspergillus oryzae

Probing the catalytically essential residues of a recombinant dipeptidyl carboxypeptidase from Escherichia coli

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