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Comparison of antioxidant properties of Teucrium polium from Golestan, North Khorasan, and Semnan Provinces
Abstracts
Introduction: Protein motions have an important role in their folding, stability and function. The proteins exhibit a variety of internal motions, that cover a wide range of amplitudes (0.01–100 A0) and time scales (1015 to > 1 s). It is generally accepted that these motions play key role in the function. Thus investigations which help to understand the relationship between stability, flexibility and function will be helpful in protein engineering and design. Living cells utilize Osmolites to increase stability of proteins without destructive effect on their function. They stabilize proteins without direct interaction with them. We wanted to know that, how the effect of sorbitol is distributed over the structure. The affected sites can play key role in the flexibility and dynamics of enzyme. Material and method: We used Hydrogen–Deuterium exchange NMR technique to study the effect of sorbitol on flexibility of hen egg white lysozyme. NMR spectra were obtained on a Bruker 500 MHZ spectrometer in the pH of 3.5 and 35 °C spectroscopy was done. Assignment of spectra was done by SPARKY software. Result: Interpretation of spectra has shown that sorbitol effect is distributed over the structure mainly in loops and alpha helices because the exchange rate in these regions decreased in the presence of sorbitol. Discussion: We found that presence of sorbitol limited the motion and dynamics of Hen Egg White Lysozyme. In this way the enzyme became stabilized. Keywords: Sorbitol, Hydrogen–Deuterium exchange NMR, Dynamics, Stability, Hen egg white lysozyme doi:10.1016/j.clinbiochem.2011.08.654
Poster – [A-10-754-1] Comparison of antioxidant properties of Teucrium polium from Golestan, North Khorasan, and Semnan Provinces Aghdas Pesaraklu, Manijeh Mianabadi, Mohammad Bagher Bagherieh- Najjar Golestan University, Faculty of Science, Department of Biology, Gorgan, Iran E-mail addresses: [email protected] (A. Pesaraklu), [email protected] (M. Mianabadi), [email protected] (M.B. Bagherieh- Najjar) Introduction: Teucrium polium (Lamiaceae) had been used for over 2000 years in traditional medicine due to its antimicrobial, antidiarrheal, anti-inflammatory, analgesic, antispasmodic, anticonvulsant and anti-diabetic properties. There are many reports on the antioxidant properties of this plant in the world. However, there is not any information about relationship between its medicinal activities and the different Iranian populations. Methods: Flowering branches of T. polium were collected from two regions in Golestan province: Ramian and Golestan Forest, two regions in North Khorasan province: Garmab and Ashkhaneh and one region in Semnan province: Shahroud. Then, they dried in dark and powdered. Antioxidant activity of their extract, phenol and flavonoid content were measured. Results: Plants (from Ramian) have the maximum of total phenol content (30.2 mg GAE/g dry weight) and plants from Garmab showed the minimum (18.06 mg GAE/g dry weight). Flavonoid content was not significantly different between Ramian and Ashkhaneh plant samples (p< 0.05) but in Shahroud was less than others. All samples had powerful ferric reducing power based on FRAP assay. The highest and the lowest reducing power had belonged to Ramian and Garmab samples, respectively. Discussion: The results suggested that T. polium from these regions is a rich source of antioxidant compounds. Nevertheless, T. polium from Ramian and Ashkhaneh have the highest antioxidant potency.
S261
Keywords: Teucrium polium, Antioxidant activity, Plant Iranian populations doi:10.1016/j.clinbiochem.2011.08.655
Poster – [A-10-755-1] Influence of Hydrophobic amino acid side groups on the acidity of aromatic imidazole ring of Histidine: A theoretical study Mohammad Reza Housaindokht, Reza Izadi, Mohammad Sadegh Sadeghi Googheri, Mohsen Sargolzaei Ferdowsi University of Mashhhad, Azadi Square, Mashhad, Iran E-mail addresses: [email protected] (M.S. Sadeghi Googheri), [email protected] (M. Sargolzaei) In this study we have calculated the acidity constant (pKa) of imidazole ring in Histidine-Hydrophobic amino acid dipeptides using the quantum chemistry and continuum solvation methods. Density functional theory (DFT) calculations with the large basis sets are used to determine the Gibbs free energy of deprotonate in the gas and liquid phases. Based on our results ΔGS values are located between − 69.38 and −18.82 kcal mol− 1 which are related to His + − Gly and His forms respectively. Aqueous-phase pKa of the dipeptides was obtained from the calculated gas-phase and solvation free energies via a thermodynamic cycle and the solvation model chemistry of Martin Karplus et al. Solvation effects are treated using a self-consistent reaction field (SCRF) formalism involving polarized continuum models (PCM). According to our calculations pKa values are between 5.50 and 8.19 that belong to His + − ILe and His + − Ala forms respectively. NBO analysis of dipeptides reveals that the electron delocalization in imidazole ring is the most effective factor in determination of acidity order at these compounds. Based on structural and atoms in molecules (AIM) analysis confirmed that the orientation of carbonyl group with respect to imidazole ring is an effective factor in imidazole ring stability. Keywords: Histidine, Acidity constant, Ab initio, Solvation free energy, DFT, AIM, NBO doi:10.1016/j.clinbiochem.2011.08.656
Poster – [A-10-764-2] Preparation and labeling of antimicrobial peptide with technetium 99mTc for infection imaging Mostafa Erfani, Mohammad Mazidi, Seyed Hassan Mirfallah, Hossein Talebi Tehran, Iran E-mail addresses: [email protected] (M. Erfani), [email protected] (M. Mazidi), [email protected] (S.H. Mirfallah), [email protected] (H. Talebi) Introduction: UBI 29–41 (a derivative of antimicrobial peptide ubiquicidin) labelled with 99mTc is reported to discriminate between bacterial infections and sterile inflammatory processes. In this study we prepared three lyophilized kits, one of them based on the direct labelling with only SnCl2 as reducing agents and two based on 6hydrazinopyridine-3-carboxylic acid (HYNIC) and tricine as coligands with or without ethylenediaminodiaceticacid (EDDA). Materials and methods: Synthesis was performed on solid phase using a standard Fmoc strategy. BOC-HYNIC was conjugated with peptide in solution. Three lyophilized kits were prepared as follows: kit 1: 20 μg UBI 29–41, 5 μg SnCl2, pH=9; kit 2: 20 μg UBI 29–41, 20 μg SnCl2, 15 mg
Introduction: Protein motions have an important role in their folding, stability and function. The proteins exhibit a variety of internal motions, that cover a wide range of amplitudes (0.01–100 A0) and time scales (1015 to > 1 s). It is generally accepted that these motions play key role in the function. Thus investigations which help to understand the relationship between stability, flexibility and function will be helpful in protein engineering and design. Living cells utilize Osmolites to increase stability of proteins without destructive effect on their function. They stabilize proteins without direct interaction with them. We wanted to know that, how the effect of sorbitol is distributed over the structure. The affected sites can play key role in the flexibility and dynamics of enzyme. Material and method: We used Hydrogen–Deuterium exchange NMR technique to study the effect of sorbitol on flexibility of hen egg white lysozyme. NMR spectra were obtained on a Bruker 500 MHZ spectrometer in the pH of 3.5 and 35 °C spectroscopy was done. Assignment of spectra was done by SPARKY software. Result: Interpretation of spectra has shown that sorbitol effect is distributed over the structure mainly in loops and alpha helices because the exchange rate in these regions decreased in the presence of sorbitol. Discussion: We found that presence of sorbitol limited the motion and dynamics of Hen Egg White Lysozyme. In this way the enzyme became stabilized. Keywords: Sorbitol, Hydrogen–Deuterium exchange NMR, Dynamics, Stability, Hen egg white lysozyme doi:10.1016/j.clinbiochem.2011.08.654
Poster – [A-10-754-1] Comparison of antioxidant properties of Teucrium polium from Golestan, North Khorasan, and Semnan Provinces Aghdas Pesaraklu, Manijeh Mianabadi, Mohammad Bagher Bagherieh- Najjar Golestan University, Faculty of Science, Department of Biology, Gorgan, Iran E-mail addresses: [email protected] (A. Pesaraklu), [email protected] (M. Mianabadi), [email protected] (M.B. Bagherieh- Najjar) Introduction: Teucrium polium (Lamiaceae) had been used for over 2000 years in traditional medicine due to its antimicrobial, antidiarrheal, anti-inflammatory, analgesic, antispasmodic, anticonvulsant and anti-diabetic properties. There are many reports on the antioxidant properties of this plant in the world. However, there is not any information about relationship between its medicinal activities and the different Iranian populations. Methods: Flowering branches of T. polium were collected from two regions in Golestan province: Ramian and Golestan Forest, two regions in North Khorasan province: Garmab and Ashkhaneh and one region in Semnan province: Shahroud. Then, they dried in dark and powdered. Antioxidant activity of their extract, phenol and flavonoid content were measured. Results: Plants (from Ramian) have the maximum of total phenol content (30.2 mg GAE/g dry weight) and plants from Garmab showed the minimum (18.06 mg GAE/g dry weight). Flavonoid content was not significantly different between Ramian and Ashkhaneh plant samples (p< 0.05) but in Shahroud was less than others. All samples had powerful ferric reducing power based on FRAP assay. The highest and the lowest reducing power had belonged to Ramian and Garmab samples, respectively. Discussion: The results suggested that T. polium from these regions is a rich source of antioxidant compounds. Nevertheless, T. polium from Ramian and Ashkhaneh have the highest antioxidant potency.
S261
Keywords: Teucrium polium, Antioxidant activity, Plant Iranian populations doi:10.1016/j.clinbiochem.2011.08.655
Poster – [A-10-755-1] Influence of Hydrophobic amino acid side groups on the acidity of aromatic imidazole ring of Histidine: A theoretical study Mohammad Reza Housaindokht, Reza Izadi, Mohammad Sadegh Sadeghi Googheri, Mohsen Sargolzaei Ferdowsi University of Mashhhad, Azadi Square, Mashhad, Iran E-mail addresses: [email protected] (M.S. Sadeghi Googheri), [email protected] (M. Sargolzaei) In this study we have calculated the acidity constant (pKa) of imidazole ring in Histidine-Hydrophobic amino acid dipeptides using the quantum chemistry and continuum solvation methods. Density functional theory (DFT) calculations with the large basis sets are used to determine the Gibbs free energy of deprotonate in the gas and liquid phases. Based on our results ΔGS values are located between − 69.38 and −18.82 kcal mol− 1 which are related to His + − Gly and His forms respectively. Aqueous-phase pKa of the dipeptides was obtained from the calculated gas-phase and solvation free energies via a thermodynamic cycle and the solvation model chemistry of Martin Karplus et al. Solvation effects are treated using a self-consistent reaction field (SCRF) formalism involving polarized continuum models (PCM). According to our calculations pKa values are between 5.50 and 8.19 that belong to His + − ILe and His + − Ala forms respectively. NBO analysis of dipeptides reveals that the electron delocalization in imidazole ring is the most effective factor in determination of acidity order at these compounds. Based on structural and atoms in molecules (AIM) analysis confirmed that the orientation of carbonyl group with respect to imidazole ring is an effective factor in imidazole ring stability. Keywords: Histidine, Acidity constant, Ab initio, Solvation free energy, DFT, AIM, NBO doi:10.1016/j.clinbiochem.2011.08.656
Poster – [A-10-764-2] Preparation and labeling of antimicrobial peptide with technetium 99mTc for infection imaging Mostafa Erfani, Mohammad Mazidi, Seyed Hassan Mirfallah, Hossein Talebi Tehran, Iran E-mail addresses: [email protected] (M. Erfani), [email protected] (M. Mazidi), [email protected] (S.H. Mirfallah), [email protected] (H. Talebi) Introduction: UBI 29–41 (a derivative of antimicrobial peptide ubiquicidin) labelled with 99mTc is reported to discriminate between bacterial infections and sterile inflammatory processes. In this study we prepared three lyophilized kits, one of them based on the direct labelling with only SnCl2 as reducing agents and two based on 6hydrazinopyridine-3-carboxylic acid (HYNIC) and tricine as coligands with or without ethylenediaminodiaceticacid (EDDA). Materials and methods: Synthesis was performed on solid phase using a standard Fmoc strategy. BOC-HYNIC was conjugated with peptide in solution. Three lyophilized kits were prepared as follows: kit 1: 20 μg UBI 29–41, 5 μg SnCl2, pH=9; kit 2: 20 μg UBI 29–41, 20 μg SnCl2, 15 mg