The influence of Al3+ion on porcine pepsin activity in vitro
Apstrakt
The in vitro effect of Al3+ ions in the concentration range 1.710-6M-8.710-3M on pepsin activity at pH 2, via kinetic parameters and its electrophoretic mobility was evaluated. Kinetic study demonstrated the existence of an activation effect of Al3+ at pH 2 on pepsin molecule. Kinetic analysis with respect to concentrations of haemoglobin showed that Al3+ ions increase the maximal velocity (Vmax) and kcat values rather than apparent affinity for substrate (KS) implying the non-competitive nature of activation which indicated that aluminium was a non-essential activator of partial non-competitive type. The values of the equilibrium constants KS and KmA for dissociation of corresponding complexes were evaluated as 0.9040.083mM and 8.560.51M, respectively. Dissociation constant KA, of activator from enzyme-activator complex calculated via kinetic and direct measurement of Al3+ binding data, as well as activation constant A50, the activator concentration that gives a rate equal to half at ...a saturating concentration of activator, were found to be 8.820.90M, 8.390.76M, and 8.050.48M respectively. Native PAGE electrophoresis shows the decrease in electrophoretic mobility of pepsin and confirms modification of the electric charge and conformational changes of pepsin caused by bound Al3+ on the pepsin molecule. Al3+ induced conformational changes of pepsin were verified by UV-VIS and IR spectra. Moreover, the absence of conformational changes in the haemoglobin molecule in the presence of Al3+ ions confirms that the obtained activation is a consequence of conformational changes caused only in the pepsin molecule.
Ključne reči:
Pepsin / aluminium / kinetics / activation / electrophoretic mobilityIzvor:
Journal of Enzyme Inhibition and Medicinal Chemistry, 2008, 23, 6, 1002-1010Finansiranje / projekti:
DOI: 10.1080/14756360701841095
ISSN: 1475-6366
PubMed: 19005946
WoS: 000260847500032
Scopus: 2-s2.0-56449095468
Kolekcije
Institucija/grupa
VinčaTY - JOUR AU - Pavelkić, Vesna M. AU - Gopčević, Kristina AU - Krstić, Danijela Z. AU - Ilić Marija A. PY - 2008 UR - https://vinar.vin.bg.ac.rs/handle/123456789/3566 AB - The in vitro effect of Al3+ ions in the concentration range 1.710-6M-8.710-3M on pepsin activity at pH 2, via kinetic parameters and its electrophoretic mobility was evaluated. Kinetic study demonstrated the existence of an activation effect of Al3+ at pH 2 on pepsin molecule. Kinetic analysis with respect to concentrations of haemoglobin showed that Al3+ ions increase the maximal velocity (Vmax) and kcat values rather than apparent affinity for substrate (KS) implying the non-competitive nature of activation which indicated that aluminium was a non-essential activator of partial non-competitive type. The values of the equilibrium constants KS and KmA for dissociation of corresponding complexes were evaluated as 0.9040.083mM and 8.560.51M, respectively. Dissociation constant KA, of activator from enzyme-activator complex calculated via kinetic and direct measurement of Al3+ binding data, as well as activation constant A50, the activator concentration that gives a rate equal to half at a saturating concentration of activator, were found to be 8.820.90M, 8.390.76M, and 8.050.48M respectively. Native PAGE electrophoresis shows the decrease in electrophoretic mobility of pepsin and confirms modification of the electric charge and conformational changes of pepsin caused by bound Al3+ on the pepsin molecule. Al3+ induced conformational changes of pepsin were verified by UV-VIS and IR spectra. Moreover, the absence of conformational changes in the haemoglobin molecule in the presence of Al3+ ions confirms that the obtained activation is a consequence of conformational changes caused only in the pepsin molecule. T2 - Journal of Enzyme Inhibition and Medicinal Chemistry T1 - The influence of Al3+ion on porcine pepsin activity in vitro VL - 23 IS - 6 SP - 1002 EP - 1010 DO - 10.1080/14756360701841095 ER -
@article{ author = "Pavelkić, Vesna M. and Gopčević, Kristina and Krstić, Danijela Z. and Ilić Marija A. ", year = "2008", abstract = "The in vitro effect of Al3+ ions in the concentration range 1.710-6M-8.710-3M on pepsin activity at pH 2, via kinetic parameters and its electrophoretic mobility was evaluated. Kinetic study demonstrated the existence of an activation effect of Al3+ at pH 2 on pepsin molecule. Kinetic analysis with respect to concentrations of haemoglobin showed that Al3+ ions increase the maximal velocity (Vmax) and kcat values rather than apparent affinity for substrate (KS) implying the non-competitive nature of activation which indicated that aluminium was a non-essential activator of partial non-competitive type. The values of the equilibrium constants KS and KmA for dissociation of corresponding complexes were evaluated as 0.9040.083mM and 8.560.51M, respectively. Dissociation constant KA, of activator from enzyme-activator complex calculated via kinetic and direct measurement of Al3+ binding data, as well as activation constant A50, the activator concentration that gives a rate equal to half at a saturating concentration of activator, were found to be 8.820.90M, 8.390.76M, and 8.050.48M respectively. Native PAGE electrophoresis shows the decrease in electrophoretic mobility of pepsin and confirms modification of the electric charge and conformational changes of pepsin caused by bound Al3+ on the pepsin molecule. Al3+ induced conformational changes of pepsin were verified by UV-VIS and IR spectra. Moreover, the absence of conformational changes in the haemoglobin molecule in the presence of Al3+ ions confirms that the obtained activation is a consequence of conformational changes caused only in the pepsin molecule.", journal = "Journal of Enzyme Inhibition and Medicinal Chemistry", title = "The influence of Al3+ion on porcine pepsin activity in vitro", volume = "23", number = "6", pages = "1002-1010", doi = "10.1080/14756360701841095" }
Pavelkić, V. M., Gopčević, K., Krstić, D. Z.,& Ilić Marija A. . (2008). The influence of Al3+ion on porcine pepsin activity in vitro. in Journal of Enzyme Inhibition and Medicinal Chemistry, 23(6), 1002-1010. https://doi.org/10.1080/14756360701841095
Pavelkić VM, Gopčević K, Krstić DZ, Ilić Marija A. . The influence of Al3+ion on porcine pepsin activity in vitro. in Journal of Enzyme Inhibition and Medicinal Chemistry. 2008;23(6):1002-1010. doi:10.1080/14756360701841095 .
Pavelkić, Vesna M., Gopčević, Kristina, Krstić, Danijela Z., Ilić Marija A. , "The influence of Al3+ion on porcine pepsin activity in vitro" in Journal of Enzyme Inhibition and Medicinal Chemistry, 23, no. 6 (2008):1002-1010, https://doi.org/10.1080/14756360701841095 . .