Structural analysis of peptide fragments following the hydrolysis of bovine serum albumin by trypsin and chymotrypsin

Özyiğit, İbrahim Ethem; Akten, Ebru Demet; Pekcan, Önder

doi:10.1080/07391102.2015.1068712

Cited by 8 publications

(6 citation statements)

References 22 publications

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“…The effect of initial BSA (1, 0.5, and 0.1 mg/mL) and TRY (2, 1, 0.5, and 0.1 mg/mL) concentrations on the reaction products were followed by analyzing the reaction medium with SDS‐PAGE. Since BSA is a protein containing lysine (59) and arginine (23) residues, hydrolysis of BSA with TRY will generate peptides with either Lys or Arg as the end group. The hydrolysis products when TRY used in free form is illustrated in Figure (a).…”

Section: Resultsmentioning

confidence: 99%

Layer‐by‐layer self‐assembly of multifunctional enzymatic UF membranes

Yürekli

2019

J of Applied Polymer Sci

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In order to eliminate membrane fouling and to ensure enzymatic hydrolysis of urea, multifunctional biocatalytic membranes were prepared by using urease (URE) and trypsin (TRY) enzymes on the sulfonated polysulfone (SPSf) ultrafiltration membrane via layer‐by‐layer (LbL) self‐assembly method. The membrane architecture consisted of multilayer assembly with TRY and URE enzymes as the outer layer and inner sandwiched layer, respectively. Polyethyleneimine (PEI) and alginate (ALG) were used as cationic and anionic polyelectrolytes. Sulfonation and PEI deposition were successfully accomplished as confirmed by attenuated total reflectance Fourier transform infrared spectroscopy, differential scanning calorimetry (DSC) and scanning electron microscopy analysis, contact angle measurements, staining with toluidine blue and Congo red dyes and dead‐end filtration experiments. A characteristic value of SPSf membrane with a high water permeability (1000 L/m2.h.bar) and 95% bovine serum albumin (BSA) rejection was observed. In static conditions, URE activities of SPSf2‐PEI‐URE membrane were not affected by BSA fouling, while TRY immobilizations with increased concentrations (SPSf2‐PEI‐URE‐PEI‐ALG‐TRY) significantly lowered the activity of URE. In dynamic conditions, each deposited layer exhibited individual resistance to flow that can be considered as irreversible fouling and caused 90% of flux decline for the SPSf2‐PEI‐URE‐PEI‐ALG‐TRY membrane assembly. The recovery of the initial flux for the multilayered membrane at the end of six fouling and washing cycles was observed 85%. Moreover, at the end of 5 cycles, 78% of the URE initial activity of the multilayered membrane was preserved. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020, 137, 48750.

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Section: Resultsmentioning

confidence: 99%

Layer‐by‐layer self‐assembly of multifunctional enzymatic UF membranes

Yürekli

2019

J of Applied Polymer Sci

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“…In addition, the presence of d ‐amino acids may increase the resistance to proteolytic degradation of the peptide while that the susceptibility of peptides to treatment with chymotrypsin is possibly related to the presence of hydrophobic residues because this enzyme cuts the peptide bond on the carboxyl side of the hydrophobic amino acid generating fragments with the hydrophobic residue at the C‐terminal end (Fernández‐Oliva et al, 2012). The degradation products formed by chymotrypsin treatment exhibited a greater retention time than the whole peptide, possibly because the fragments usually form a cluster, which creates a more hydrophobic environment (Özyiǧit et al, 2016).…”

Section: Resultsmentioning

confidence: 99%

“…It may be due to a degradation product, suggesting that the peptide has low susceptibility to trypsin treatment (Figure 2b, left, blue and red). This signal probably is related to the appearance of some excimer (Özyiǧit et al, 2016) associated with peptide cleavage at arginine, which generates a more hydrophobic fragment that has a greater t R .…”

Section: Resultsmentioning

confidence: 99%

In‐house standards derived from doping peptides: Enzymatic and serum stability and degradation profile of GHRP and GHRH‐related peptides

González‐López,

Guerra‐Acero‐Turizo,

Blanco‐Medina

et al. 2023

Biomedical Chromatography

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Matrix effect and sample pretreatment significantly affect the percentage recovery of peptides in biological matrices, affecting the method robustness and accuracy. To counteract this effect, an internal standard (IS) is used; however, in most cases this is not available, which limits the analytical method. It is important to identify short peptides that can be used as ISs in the quantification of peptides in biological matrices. In this study, doping peptides GHRP‐4, GHRP‐5, GHRP‐6, Sermorelin (1–11), Sermorelin (13–20) and Sermorelin (22–29) were synthesized using solid‐phase peptide synthesis. Treatment with human blood, trypsin and chymotrypsin was used to determine the stability of the peptides. Products were evaluated using the high‐performance liquid chromatography‐diode array detector (HPLC‐DAD) method. The analytical methodology and sample pretreatment were effective for the analysis of these molecules. A unique profile related to protein binding and enzymatic stability of each peptide was established. GHRP‐4, GHRP‐6 and Sermorelin (22–29) can be considered as in‐house ISs as they were stable to enzyme and blood treatment and can be used for the quantification of peptides in biological samples. Peptides GHRP‐6 and Sermorelin (22–29) were used to analyse a dimeric peptide (26[F] LfcinB (20–30)2) in four different matrices to test these peptides as in‐house IS.

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“…Time‐resolved fluorescence technology can more accurately distinguish the quenching mechanism between ligand and the protein . [ 43 ] Therefore, we measured the excited state lifetime of CAPE and protease to further determine the quenching mechanism of protease with CAPE. A fluorescence decay curve was fitted based on double exponential fitting, and the average life span of protease was calculated using the following formula:

τ goodbreak= α_{1} \cdot τ_{1} goodbreak+ α_{2} \cdot τ_{2}

where

τ

is the fluorescence lifetime of protease,

τ_{1}

and

τ_{2}

are decay times, and

α_{1}

α_{2}

are pre‐exponential factors.…”

Section: Resultsmentioning

confidence: 99%

Interaction between caffeic acid phenethyl ester and protease: monitoring by spectroscopic and molecular docking approaches

et al. 2022

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The interaction of one anticancer drug (caffeic acid phenethyl ester; CAPE) with three proteases (trypsin, pepsin and α-chymotrypsin) has been investigated with multispectral methods and molecular docking. As an active components in propolis, the findings are of great benefit to metabolism, design, and structural modification of drugs. The results show that CAPE has an obvious ability to quench the trypsin, pepsin, or α-chymotrypsin fluorescence mainly through a static quenching procedure.Trypsin has the largest binding affinity to CAPE, and α-chymotrypsin has the smallest binding affinity to CAPE. The data obtained from thermodynamic parameters and molecular docking prove that the spontaneously interaction between CAPE and each protease is mainly due to a combination of van der Waals (vdW) force and hydrogen bond (H-bond), controlled by an enthalpy-driven process. The binding force, strength, position, and the number of H-bond are further obtained from the results of molecular docking. Through ultraviolet spectroscopy, dynamic light scattering and circular dichroism experiments, the change in the protease secondary structure induced by CAPE was observed. Additionally, the addition of protease had a positive effect on the antioxidative activity of CAPE, and α-chymotrypsin has the greatest effect on the removal of 2,2-diphenyl-1-picrylhydrazyl free radicals by CAPE.α-chymotrypsin, caffeic acid phenethyl ester (CAPE), pepsin, trypsin | INTRODUCTIONCaffeic acid phenethyl ester (CAPE) has been widely used in various regions as a traditional natural medicine. [1] As the main active ingredient of propolis, CAPE has various biological activities, including anticancer, anti-inflammatory, antibacterial, neuroprotective, etc. [2][3][4] In addition, it has a very strong antioxidant capacity, [5] as a polyphenol containing hydroxyl groups in benzenediol. [6] In previous works, the study of the interaction between CAPE and macromolecules mainly focus on the binding mechanism, influencing factors, drug activities, and pharmacology; many theoretical researches have been done.Studies have shown that poly(3-hydroxybutyrate) (PHB) fibre can enhance the antioxidant and antibacterial activities of CAPE. [7] CAPE can inhibit iNOS gene transcription by inhibiting the nuclear factor NF-κB site, and can inhibit the catalytic activity of iNOS, [8] which provides a research basis for the anti-inflammatory and antitumour effects of CAPE. According to reports, after CAPE is compounded with γ-cyclodextrin (γ-CD), the anticancer activity of CAPE will increase. [9] In addition, in the work of this research group, the binding constant of CAPE and bovine serum albumin was $10 6 , of which the main forces are H-bond and vdW force, and the driving mode is enthalpy drive. However, there are few studies on the interaction

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Structural analysis of peptide fragments following the hydrolysis of bovine serum albumin by trypsin and chymotrypsin

Cited by 8 publications

References 22 publications

Layer‐by‐layer self‐assembly of multifunctional enzymatic UF membranes

Layer‐by‐layer self‐assembly of multifunctional enzymatic UF membranes

In‐house standards derived from doping peptides: Enzymatic and serum stability and degradation profile of GHRP and GHRH‐related peptides

Interaction between caffeic acid phenethyl ester and protease: monitoring by spectroscopic and molecular docking approaches

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