TY  - JOUR
AU  - Grozdanović, Milica M.
AU  - Čavić, Milena
AU  - Nešić, Andrijana N.
AU  - Anđelković, Uroš
AU  - Akbari, Peyman
AU  - Smit, Joost
AU  - Gavrović-Jankulović, Marija
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/2039
AB  - Background: The intestinal epithelium forms a barrier that food allergens must cross in order to induce sensitization. The aim of this study was to evaluate the impact of the plant-derived food cysteine protease - actinidin (Act d1) on the integrity of intestinal epithelium tight junctions (TJs). Methods: Effects of Act d1 on the intestinal epithelium were evaluated in Caco-2 monolayers and in a mouse model by measuring transepithelial resistance and in vivo permeability. Integrity of the tight junctions was analyzed by confocal microscopy. Proteolysis of TJ protein occludin was evaluated by mass spectrometry. Results: Actinidin (1 mg/mL) reduced the transepithelial resistance of the cell monolayer by 18.1% (after 1 h) and 25.6% (after 4 h). This loss of barrier function was associated with Act d 1 disruption of the occludin and zonula occludens (ZO)-1 network. The effect on intestinal permeability in vivo was demonstrated by the significantly higher concentration of 40 kDa FITC-dextran (233 mu g/mL) that passed from the intestine into the serum of Act d1 treated mice in comparison to the control group (0.5 mu g/mL). Human occludin was fragmented, and putative Act d1 cleavage sites were identified in extracellular loops of human occludin. Conclusion: Act d1 caused protease-dependent disruption of tight junctions in confluent Caco-2 cells and increased intestinal permeability in mice. General significance: In line with the observed effects of food cysteine proteases in occupational allergy, these results suggest that disruption of tight junctions by food cysteine proteases may contribute to the process of sensitization in food allergy. (C) 2015 Elsevier B.V. All rights reserved.
PB  - Elsevier Science Bv, Amsterdam
T2  - Biochimica et Biophysica Acta: General Subjects
T1  - Kiwifruit cysteine protease actinidin compromises the intestinal barrier by disrupting tight junctions
VL  - 1860
IS  - 3
SP  - 516
EP  - 526
DO  - 10.1016/j.bbagen.2015.12.005
ER  - 

@article{
author = "Grozdanović, Milica M. and Čavić, Milena and Nešić, Andrijana N. and Anđelković, Uroš and Akbari, Peyman and Smit, Joost and Gavrović-Jankulović, Marija",
year = "2016",
abstract = "Background: The intestinal epithelium forms a barrier that food allergens must cross in order to induce sensitization. The aim of this study was to evaluate the impact of the plant-derived food cysteine protease - actinidin (Act d1) on the integrity of intestinal epithelium tight junctions (TJs). Methods: Effects of Act d1 on the intestinal epithelium were evaluated in Caco-2 monolayers and in a mouse model by measuring transepithelial resistance and in vivo permeability. Integrity of the tight junctions was analyzed by confocal microscopy. Proteolysis of TJ protein occludin was evaluated by mass spectrometry. Results: Actinidin (1 mg/mL) reduced the transepithelial resistance of the cell monolayer by 18.1% (after 1 h) and 25.6% (after 4 h). This loss of barrier function was associated with Act d 1 disruption of the occludin and zonula occludens (ZO)-1 network. The effect on intestinal permeability in vivo was demonstrated by the significantly higher concentration of 40 kDa FITC-dextran (233 mu g/mL) that passed from the intestine into the serum of Act d1 treated mice in comparison to the control group (0.5 mu g/mL). Human occludin was fragmented, and putative Act d1 cleavage sites were identified in extracellular loops of human occludin. Conclusion: Act d1 caused protease-dependent disruption of tight junctions in confluent Caco-2 cells and increased intestinal permeability in mice. General significance: In line with the observed effects of food cysteine proteases in occupational allergy, these results suggest that disruption of tight junctions by food cysteine proteases may contribute to the process of sensitization in food allergy. (C) 2015 Elsevier B.V. All rights reserved.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Biochimica et Biophysica Acta: General Subjects",
title = "Kiwifruit cysteine protease actinidin compromises the intestinal barrier by disrupting tight junctions",
volume = "1860",
number = "3",
pages = "516-526",
doi = "10.1016/j.bbagen.2015.12.005"
}

Grozdanović, M. M., Čavić, M., Nešić, A. N., Anđelković, U., Akbari, P., Smit, J.,& Gavrović-Jankulović, M.. (2016). Kiwifruit cysteine protease actinidin compromises the intestinal barrier by disrupting tight junctions. in Biochimica et Biophysica Acta: General Subjects
Elsevier Science Bv, Amsterdam., 1860(3), 516-526.
https://doi.org/10.1016/j.bbagen.2015.12.005

Grozdanović MM, Čavić M, Nešić AN, Anđelković U, Akbari P, Smit J, Gavrović-Jankulović M. Kiwifruit cysteine protease actinidin compromises the intestinal barrier by disrupting tight junctions. in Biochimica et Biophysica Acta: General Subjects. 2016;1860(3):516-526.
doi:10.1016/j.bbagen.2015.12.005 .

Grozdanović, Milica M., Čavić, Milena, Nešić, Andrijana N., Anđelković, Uroš, Akbari, Peyman, Smit, Joost, Gavrović-Jankulović, Marija, "Kiwifruit cysteine protease actinidin compromises the intestinal barrier by disrupting tight junctions" in Biochimica et Biophysica Acta: General Subjects, 1860, no. 3 (2016):516-526,
https://doi.org/10.1016/j.bbagen.2015.12.005 . .

TY  - JOUR
AU  - Grozdanović, Milica M.
AU  - Čavić, Milena
AU  - Nešić, Andrijana N.
AU  - Anđelković, Uroš
AU  - Akbari, Peyman
AU  - Smit, Joost
AU  - Gavrović-Jankulović, Marija
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3559
AB  - Background: The intestinal epithelium forms a barrier that food allergens must cross in order to induce sensitization. The aim of this study was to evaluate the impact of the plant-derived food cysteine protease - actinidin (Act d1) on the integrity of intestinal epithelium tight junctions (TJs). Methods: Effects of Act d1 on the intestinal epithelium were evaluated in Caco-2 monolayers and in a mouse model by measuring transepithelial resistance and in vivo permeability. Integrity of the tight junctions was analyzed by confocal microscopy. Proteolysis of TJ protein occludin was evaluated by mass spectrometry. Results: Actinidin (1 mg/mL) reduced the transepithelial resistance of the cell monolayer by 18.1% (after 1 h) and 25.6% (after 4 h). This loss of barrier function was associated with Act d 1 disruption of the occludin and zonula occludens (ZO)-1 network. The effect on intestinal permeability in vivo was demonstrated by the significantly higher concentration of 40 kDa FITC-dextran (233 mu g/mL) that passed from the intestine into the serum of Act d1 treated mice in comparison to the control group (0.5 mu g/mL). Human occludin was fragmented, and putative Act d1 cleavage sites were identified in extracellular loops of human occludin. Conclusion: Act d1 caused protease-dependent disruption of tight junctions in confluent Caco-2 cells and increased intestinal permeability in mice. General significance: In line with the observed effects of food cysteine proteases in occupational allergy, these results suggest that disruption of tight junctions by food cysteine proteases may contribute to the process of sensitization in food allergy. (C) 2015 Elsevier B.V. All rights reserved.
PB  - Elsevier Science Bv, Amsterdam
T2  - Biochimica et Biophysica Acta: General Subjects
T1  - Kiwifruit cysteine protease actinidin compromises the intestinal barrier by disrupting tight junctions
VL  - 1860
IS  - 3
SP  - 516
EP  - 526
DO  - 10.1016/j.bbagen.2015.12.005
ER  - 

@article{
author = "Grozdanović, Milica M. and Čavić, Milena and Nešić, Andrijana N. and Anđelković, Uroš and Akbari, Peyman and Smit, Joost and Gavrović-Jankulović, Marija",
year = "2016",
abstract = "Background: The intestinal epithelium forms a barrier that food allergens must cross in order to induce sensitization. The aim of this study was to evaluate the impact of the plant-derived food cysteine protease - actinidin (Act d1) on the integrity of intestinal epithelium tight junctions (TJs). Methods: Effects of Act d1 on the intestinal epithelium were evaluated in Caco-2 monolayers and in a mouse model by measuring transepithelial resistance and in vivo permeability. Integrity of the tight junctions was analyzed by confocal microscopy. Proteolysis of TJ protein occludin was evaluated by mass spectrometry. Results: Actinidin (1 mg/mL) reduced the transepithelial resistance of the cell monolayer by 18.1% (after 1 h) and 25.6% (after 4 h). This loss of barrier function was associated with Act d 1 disruption of the occludin and zonula occludens (ZO)-1 network. The effect on intestinal permeability in vivo was demonstrated by the significantly higher concentration of 40 kDa FITC-dextran (233 mu g/mL) that passed from the intestine into the serum of Act d1 treated mice in comparison to the control group (0.5 mu g/mL). Human occludin was fragmented, and putative Act d1 cleavage sites were identified in extracellular loops of human occludin. Conclusion: Act d1 caused protease-dependent disruption of tight junctions in confluent Caco-2 cells and increased intestinal permeability in mice. General significance: In line with the observed effects of food cysteine proteases in occupational allergy, these results suggest that disruption of tight junctions by food cysteine proteases may contribute to the process of sensitization in food allergy. (C) 2015 Elsevier B.V. All rights reserved.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Biochimica et Biophysica Acta: General Subjects",
title = "Kiwifruit cysteine protease actinidin compromises the intestinal barrier by disrupting tight junctions",
volume = "1860",
number = "3",
pages = "516-526",
doi = "10.1016/j.bbagen.2015.12.005"
}

Grozdanović, M. M., Čavić, M., Nešić, A. N., Anđelković, U., Akbari, P., Smit, J.,& Gavrović-Jankulović, M.. (2016). Kiwifruit cysteine protease actinidin compromises the intestinal barrier by disrupting tight junctions. in Biochimica et Biophysica Acta: General Subjects
Elsevier Science Bv, Amsterdam., 1860(3), 516-526.
https://doi.org/10.1016/j.bbagen.2015.12.005

Grozdanović MM, Čavić M, Nešić AN, Anđelković U, Akbari P, Smit J, Gavrović-Jankulović M. Kiwifruit cysteine protease actinidin compromises the intestinal barrier by disrupting tight junctions. in Biochimica et Biophysica Acta: General Subjects. 2016;1860(3):516-526.
doi:10.1016/j.bbagen.2015.12.005 .

Grozdanović, Milica M., Čavić, Milena, Nešić, Andrijana N., Anđelković, Uroš, Akbari, Peyman, Smit, Joost, Gavrović-Jankulović, Marija, "Kiwifruit cysteine protease actinidin compromises the intestinal barrier by disrupting tight junctions" in Biochimica et Biophysica Acta: General Subjects, 1860, no. 3 (2016):516-526,
https://doi.org/10.1016/j.bbagen.2015.12.005 . .

TY  - JOUR
AU  - Grozdanović, Milica M.
AU  - Ostojić, Sanja B.
AU  - Aleksić, Ivana
AU  - Anđelković, Uroš
AU  - Petersen, Arnd
AU  - Gavrović-Jankulović, Marija
PY  - 2014
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1859
AB  - BACKGROUND: Actinidin is a cysteine protease and major allergen from kiwi fruit. When purified under specific native conditions, actinidin preparations from fresh kiwi fruit contain both an active and inactive form of this enzyme. In this study, biochemical and immunological properties upon simulated gastro-intestinal digestion, as well as thermal stability, were investigated for both active and E-64-inhibited actinidin. RESULTS: Active actinidin retained its primary structure and proteolytic activity after 2 h of simulated gastric digestion, followed by 2 h of intestinal digestion, as assessed by SDS-PAGE, zymography and mass spectroscopy. Immunological reactivity of active actinidin was also preserved, as tested by immunoelectrophoresis. The E-64 inhibited actinidin was fully degraded after 1 h of pepsin treatment. Differential scanning calorimetry showed that active actinidin has one transition maximum temperature (T-m) at 73.9 degrees C, whereas in the E-64-actinidin complex the two actinidin domains unfolded independently, with the first domain having a T-m value of only 61 degrees C. CONCLUSION: Active actinidin is capable of reaching the intestinal mucosa in a proteolytically active and immunogenic state. Inhibitor binding induces changes in the actinidin molecule that go beyond inhibition of proteolytic activity, also influencing the digestion stability and T-m values of actinidin, features important in the characterisation of food allergens. (C) 2014 Society of Chemical Industry
PB  - Wiley-Blackwell, Hoboken
T2  - Journal of the Science of Food and Agriculture
T1  - Active actinidin retains function upon gastro-intestinal digestion and is more thermostable than the E-64-inhibited counterpart
VL  - 94
IS  - 14
SP  - 3046
EP  - 3052
DO  - 10.1002/jsfa.6656
ER  - 

@article{
author = "Grozdanović, Milica M. and Ostojić, Sanja B. and Aleksić, Ivana and Anđelković, Uroš and Petersen, Arnd and Gavrović-Jankulović, Marija",
year = "2014",
abstract = "BACKGROUND: Actinidin is a cysteine protease and major allergen from kiwi fruit. When purified under specific native conditions, actinidin preparations from fresh kiwi fruit contain both an active and inactive form of this enzyme. In this study, biochemical and immunological properties upon simulated gastro-intestinal digestion, as well as thermal stability, were investigated for both active and E-64-inhibited actinidin. RESULTS: Active actinidin retained its primary structure and proteolytic activity after 2 h of simulated gastric digestion, followed by 2 h of intestinal digestion, as assessed by SDS-PAGE, zymography and mass spectroscopy. Immunological reactivity of active actinidin was also preserved, as tested by immunoelectrophoresis. The E-64 inhibited actinidin was fully degraded after 1 h of pepsin treatment. Differential scanning calorimetry showed that active actinidin has one transition maximum temperature (T-m) at 73.9 degrees C, whereas in the E-64-actinidin complex the two actinidin domains unfolded independently, with the first domain having a T-m value of only 61 degrees C. CONCLUSION: Active actinidin is capable of reaching the intestinal mucosa in a proteolytically active and immunogenic state. Inhibitor binding induces changes in the actinidin molecule that go beyond inhibition of proteolytic activity, also influencing the digestion stability and T-m values of actinidin, features important in the characterisation of food allergens. (C) 2014 Society of Chemical Industry",
publisher = "Wiley-Blackwell, Hoboken",
journal = "Journal of the Science of Food and Agriculture",
title = "Active actinidin retains function upon gastro-intestinal digestion and is more thermostable than the E-64-inhibited counterpart",
volume = "94",
number = "14",
pages = "3046-3052",
doi = "10.1002/jsfa.6656"
}

Grozdanović, M. M., Ostojić, S. B., Aleksić, I., Anđelković, U., Petersen, A.,& Gavrović-Jankulović, M.. (2014). Active actinidin retains function upon gastro-intestinal digestion and is more thermostable than the E-64-inhibited counterpart. in Journal of the Science of Food and Agriculture
Wiley-Blackwell, Hoboken., 94(14), 3046-3052.
https://doi.org/10.1002/jsfa.6656

Grozdanović MM, Ostojić SB, Aleksić I, Anđelković U, Petersen A, Gavrović-Jankulović M. Active actinidin retains function upon gastro-intestinal digestion and is more thermostable than the E-64-inhibited counterpart. in Journal of the Science of Food and Agriculture. 2014;94(14):3046-3052.
doi:10.1002/jsfa.6656 .

Grozdanović, Milica M., Ostojić, Sanja B., Aleksić, Ivana, Anđelković, Uroš, Petersen, Arnd, Gavrović-Jankulović, Marija, "Active actinidin retains function upon gastro-intestinal digestion and is more thermostable than the E-64-inhibited counterpart" in Journal of the Science of Food and Agriculture, 94, no. 14 (2014):3046-3052,
https://doi.org/10.1002/jsfa.6656 . .

TY  - JOUR
AU  - Čavić, Milena
AU  - Grozdanović, Milica M.
AU  - Bajić, Aleksandar
AU  - Jankovic, Radmila
AU  - Andjus, Pavle R.
AU  - Gavrović-Jankulović, Marija
PY  - 2014
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1866
AB  - Actinidin, a kiwifruit cysteine protease, is a marker allergen for genuine sensitization to this food allergen source. Inhalatory cysteine proteases have the capacity for disruption of tight junctions (TJs) enhancing the permeability of the bronchial epithelium. No such properties have been reported for allergenic food proteases so far. The aim was to determine the effect of actinidin on the integrity of T84 monolayers by evaluating its action on the TJ protein occludin. Immunoblot and immunofluorescence were employed for the detection of occludin protein alterations. Gene expression was evaluated by RT-PCR. Breach of occludin network was assessed by measuring transepithelial resistance, blue dextran leakage and passage of allergens from the apical to basolateral compartment. Actinidin exerted direct proteolytic cleavage of occludin; no alteration of occludin gene expression was detected. There was a reduction of occludin staining upon actinidin treatment as a consequence of its degradation and dispersion within the membrane. There was an increase in permeability of the T84 monolayer resulting in reduced transepithelial resistance, blue dextran leakage and passage of allergens actinidin and thaumatin-like protein from the apical to basolateral compartment. Opening of TJs by actinidin may increase intestinal permeability and contribute to the process of sensitization in kiwifruit allergy. (C) 2014 Elsevier Ltd. All rights reserved.
PB  - Pergamon-Elsevier Science Ltd, Oxford
T2  - Food and Chemical Toxicology
T1  - The effect of kiwifruit (Actinidia deliciosa) cysteine protease actinidin on the occludin tight junction network in T84 intestinal epithelial cells
VL  - 72
SP  - 61
EP  - 68
DO  - 10.1016/j.fct.2014.07.012
ER  - 

@article{
author = "Čavić, Milena and Grozdanović, Milica M. and Bajić, Aleksandar and Jankovic, Radmila and Andjus, Pavle R. and Gavrović-Jankulović, Marija",
year = "2014",
abstract = "Actinidin, a kiwifruit cysteine protease, is a marker allergen for genuine sensitization to this food allergen source. Inhalatory cysteine proteases have the capacity for disruption of tight junctions (TJs) enhancing the permeability of the bronchial epithelium. No such properties have been reported for allergenic food proteases so far. The aim was to determine the effect of actinidin on the integrity of T84 monolayers by evaluating its action on the TJ protein occludin. Immunoblot and immunofluorescence were employed for the detection of occludin protein alterations. Gene expression was evaluated by RT-PCR. Breach of occludin network was assessed by measuring transepithelial resistance, blue dextran leakage and passage of allergens from the apical to basolateral compartment. Actinidin exerted direct proteolytic cleavage of occludin; no alteration of occludin gene expression was detected. There was a reduction of occludin staining upon actinidin treatment as a consequence of its degradation and dispersion within the membrane. There was an increase in permeability of the T84 monolayer resulting in reduced transepithelial resistance, blue dextran leakage and passage of allergens actinidin and thaumatin-like protein from the apical to basolateral compartment. Opening of TJs by actinidin may increase intestinal permeability and contribute to the process of sensitization in kiwifruit allergy. (C) 2014 Elsevier Ltd. All rights reserved.",
publisher = "Pergamon-Elsevier Science Ltd, Oxford",
journal = "Food and Chemical Toxicology",
title = "The effect of kiwifruit (Actinidia deliciosa) cysteine protease actinidin on the occludin tight junction network in T84 intestinal epithelial cells",
volume = "72",
pages = "61-68",
doi = "10.1016/j.fct.2014.07.012"
}

Čavić, M., Grozdanović, M. M., Bajić, A., Jankovic, R., Andjus, P. R.,& Gavrović-Jankulović, M.. (2014). The effect of kiwifruit (Actinidia deliciosa) cysteine protease actinidin on the occludin tight junction network in T84 intestinal epithelial cells. in Food and Chemical Toxicology
Pergamon-Elsevier Science Ltd, Oxford., 72, 61-68.
https://doi.org/10.1016/j.fct.2014.07.012

Čavić M, Grozdanović MM, Bajić A, Jankovic R, Andjus PR, Gavrović-Jankulović M. The effect of kiwifruit (Actinidia deliciosa) cysteine protease actinidin on the occludin tight junction network in T84 intestinal epithelial cells. in Food and Chemical Toxicology. 2014;72:61-68.
doi:10.1016/j.fct.2014.07.012 .

Čavić, Milena, Grozdanović, Milica M., Bajić, Aleksandar, Jankovic, Radmila, Andjus, Pavle R., Gavrović-Jankulović, Marija, "The effect of kiwifruit (Actinidia deliciosa) cysteine protease actinidin on the occludin tight junction network in T84 intestinal epithelial cells" in Food and Chemical Toxicology, 72 (2014):61-68,
https://doi.org/10.1016/j.fct.2014.07.012 . .

TY  - DATA
AU  - Grozdanović, Milica M.
AU  - Drakulić, Branko J.
AU  - Gavrović-Jankulović, Marija
PY  - 2013
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3476
PB  - Elsevier Science Bv, Amsterdam
T2  - Biochimica et Biophysica Acta: General Subjects
T1  - Supplementary data for article: Grozdanovic, M. M.; Drakulić, B. J.; Gavrović-Jankulović, M. Conformational Mobility of Active and E-64-Inhibited Actinidin. Biochimica et Biophysica Acta: General Subjects 2013, 1830 (10), 4790–4799. https://doi.org/10.1016/j.bbagen.2013.06.015
UR  - https://hdl.handle.net/21.15107/rcub_cherry_3476
ER  - 

@misc{
author = "Grozdanović, Milica M. and Drakulić, Branko J. and Gavrović-Jankulović, Marija",
year = "2013",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Biochimica et Biophysica Acta: General Subjects",
title = "Supplementary data for article: Grozdanovic, M. M.; Drakulić, B. J.; Gavrović-Jankulović, M. Conformational Mobility of Active and E-64-Inhibited Actinidin. Biochimica et Biophysica Acta: General Subjects 2013, 1830 (10), 4790–4799. https://doi.org/10.1016/j.bbagen.2013.06.015",
url = "https://hdl.handle.net/21.15107/rcub_cherry_3476"
}

Grozdanović, M. M., Drakulić, B. J.,& Gavrović-Jankulović, M.. (2013). Supplementary data for article: Grozdanovic, M. M.; Drakulić, B. J.; Gavrović-Jankulović, M. Conformational Mobility of Active and E-64-Inhibited Actinidin. Biochimica et Biophysica Acta: General Subjects 2013, 1830 (10), 4790–4799. https://doi.org/10.1016/j.bbagen.2013.06.015. in Biochimica et Biophysica Acta: General Subjects
Elsevier Science Bv, Amsterdam..
https://hdl.handle.net/21.15107/rcub_cherry_3476

Grozdanović MM, Drakulić BJ, Gavrović-Jankulović M. Supplementary data for article: Grozdanovic, M. M.; Drakulić, B. J.; Gavrović-Jankulović, M. Conformational Mobility of Active and E-64-Inhibited Actinidin. Biochimica et Biophysica Acta: General Subjects 2013, 1830 (10), 4790–4799. https://doi.org/10.1016/j.bbagen.2013.06.015. in Biochimica et Biophysica Acta: General Subjects. 2013;.
https://hdl.handle.net/21.15107/rcub_cherry_3476 .

Grozdanović, Milica M., Drakulić, Branko J., Gavrović-Jankulović, Marija, "Supplementary data for article: Grozdanovic, M. M.; Drakulić, B. J.; Gavrović-Jankulović, M. Conformational Mobility of Active and E-64-Inhibited Actinidin. Biochimica et Biophysica Acta: General Subjects 2013, 1830 (10), 4790–4799. https://doi.org/10.1016/j.bbagen.2013.06.015" in Biochimica et Biophysica Acta: General Subjects (2013),
https://hdl.handle.net/21.15107/rcub_cherry_3476 .

TY  - DATA
AU  - Grozdanović, Milica M.
AU  - Burazer, Lidija M.
AU  - Gavrović-Jankulović, Marija
PY  - 2013
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3502
PB  - Elsevier Sci Ltd, Oxford
T2  - International Dairy Journal
T1  - Supplementary data for article: Grozdanović, M. M.; Burazer, L. M.; Gavrović-Jankulović, M. Kiwifruit (Actinidia Deliciosa) Extract Shows Potential as a Low-Cost and Efficient Milk-Clotting Agent. International Dairy Journal 2013, 32 (1), 46–52. https://doi.org/10.1016/j.idairyj.2013.03.001
UR  - https://hdl.handle.net/21.15107/rcub_cherry_3502
ER  - 

@misc{
author = "Grozdanović, Milica M. and Burazer, Lidija M. and Gavrović-Jankulović, Marija",
year = "2013",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "International Dairy Journal",
title = "Supplementary data for article: Grozdanović, M. M.; Burazer, L. M.; Gavrović-Jankulović, M. Kiwifruit (Actinidia Deliciosa) Extract Shows Potential as a Low-Cost and Efficient Milk-Clotting Agent. International Dairy Journal 2013, 32 (1), 46–52. https://doi.org/10.1016/j.idairyj.2013.03.001",
url = "https://hdl.handle.net/21.15107/rcub_cherry_3502"
}

Grozdanović, M. M., Burazer, L. M.,& Gavrović-Jankulović, M.. (2013). Supplementary data for article: Grozdanović, M. M.; Burazer, L. M.; Gavrović-Jankulović, M. Kiwifruit (Actinidia Deliciosa) Extract Shows Potential as a Low-Cost and Efficient Milk-Clotting Agent. International Dairy Journal 2013, 32 (1), 46–52. https://doi.org/10.1016/j.idairyj.2013.03.001. in International Dairy Journal
Elsevier Sci Ltd, Oxford..
https://hdl.handle.net/21.15107/rcub_cherry_3502

Grozdanović MM, Burazer LM, Gavrović-Jankulović M. Supplementary data for article: Grozdanović, M. M.; Burazer, L. M.; Gavrović-Jankulović, M. Kiwifruit (Actinidia Deliciosa) Extract Shows Potential as a Low-Cost and Efficient Milk-Clotting Agent. International Dairy Journal 2013, 32 (1), 46–52. https://doi.org/10.1016/j.idairyj.2013.03.001. in International Dairy Journal. 2013;.
https://hdl.handle.net/21.15107/rcub_cherry_3502 .

Grozdanović, Milica M., Burazer, Lidija M., Gavrović-Jankulović, Marija, "Supplementary data for article: Grozdanović, M. M.; Burazer, L. M.; Gavrović-Jankulović, M. Kiwifruit (Actinidia Deliciosa) Extract Shows Potential as a Low-Cost and Efficient Milk-Clotting Agent. International Dairy Journal 2013, 32 (1), 46–52. https://doi.org/10.1016/j.idairyj.2013.03.001" in International Dairy Journal (2013),
https://hdl.handle.net/21.15107/rcub_cherry_3502 .

TY  - JOUR
AU  - Grozdanović, Milica M.
AU  - Burazer, Lidija M.
AU  - Gavrović-Jankulović, Marija
PY  - 2013
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1589
AB  - Actinidin, a cysteine protease accounting for more than 50% of the soluble proteins in kiwifruit pulp, has shown promise as a milk-clotting agent. In this study, the potential use of kiwifruit pulp extract as a clotting agent was investigated. It was shown that three kiwifruit extracts made from the pulp of a single fruit have significantly different levels of active actinidin, depending on the extraction buffer employed. Kiwifruit extract prepared at pH 5.0 had the best milk-clotting properties, with a nearly 30% better ratio of clotting activity to proteolytic activity than purified actinidin. This extract produced a casein coagulum clearly separated from the whey proteins, and was shown to be stable at room temperature for up to two months. This extract has the potential to be employed as an efficient and low-cost milk-clotting agent in the production of dairy products.
PB  - Elsevier Sci Ltd, Oxford
T2  - International Dairy Journal
T1  - Kiwifruit (Actinidia deliciosa) extract shows potential as a low-cost and efficient milk-clotting agent
VL  - 32
IS  - 1
SP  - 46
EP  - 52
DO  - 10.1016/j.idairyj.2013.03.001
ER  - 

@article{
author = "Grozdanović, Milica M. and Burazer, Lidija M. and Gavrović-Jankulović, Marija",
year = "2013",
abstract = "Actinidin, a cysteine protease accounting for more than 50% of the soluble proteins in kiwifruit pulp, has shown promise as a milk-clotting agent. In this study, the potential use of kiwifruit pulp extract as a clotting agent was investigated. It was shown that three kiwifruit extracts made from the pulp of a single fruit have significantly different levels of active actinidin, depending on the extraction buffer employed. Kiwifruit extract prepared at pH 5.0 had the best milk-clotting properties, with a nearly 30% better ratio of clotting activity to proteolytic activity than purified actinidin. This extract produced a casein coagulum clearly separated from the whey proteins, and was shown to be stable at room temperature for up to two months. This extract has the potential to be employed as an efficient and low-cost milk-clotting agent in the production of dairy products.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "International Dairy Journal",
title = "Kiwifruit (Actinidia deliciosa) extract shows potential as a low-cost and efficient milk-clotting agent",
volume = "32",
number = "1",
pages = "46-52",
doi = "10.1016/j.idairyj.2013.03.001"
}

Grozdanović, M. M., Burazer, L. M.,& Gavrović-Jankulović, M.. (2013). Kiwifruit (Actinidia deliciosa) extract shows potential as a low-cost and efficient milk-clotting agent. in International Dairy Journal
Elsevier Sci Ltd, Oxford., 32(1), 46-52.
https://doi.org/10.1016/j.idairyj.2013.03.001

Grozdanović MM, Burazer LM, Gavrović-Jankulović M. Kiwifruit (Actinidia deliciosa) extract shows potential as a low-cost and efficient milk-clotting agent. in International Dairy Journal. 2013;32(1):46-52.
doi:10.1016/j.idairyj.2013.03.001 .

Grozdanović, Milica M., Burazer, Lidija M., Gavrović-Jankulović, Marija, "Kiwifruit (Actinidia deliciosa) extract shows potential as a low-cost and efficient milk-clotting agent" in International Dairy Journal, 32, no. 1 (2013):46-52,
https://doi.org/10.1016/j.idairyj.2013.03.001 . .

TY  - JOUR
AU  - Macinkovic, Igor S.
AU  - Abughren, Mohamed
AU  - Mrkić, Ivan
AU  - Grozdanović, Milica M.
AU  - Prodanović, Radivoje
AU  - Gavrović-Jankulović, Marija
PY  - 2013
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1449
AB  - High levels of recombinant protein expression can lead to the formation of insoluble inclusion bodies. These complex aggregates are commonly solubilized in strong denaturants, such as 6-8 M urea, although, if possible, solubilization under milder conditions could facilitate subsequent refolding and purification of bioactive proteins. Commercially available GST-tag assays are designed for quantitative measurement of GST activity under native conditions. GST fusion proteins accumulated in inclusion bodies are considered to be undetectable by such assays. In this work, solubilization of recombinantly produced proteins was performed in 4 M urea. The activity of rGST was assayed in 2 M urea and it was shown that rGST preserves 85% of its activity under such denaturing conditions. A colorimetric GST activity assay with 1-chloro-2, 4-dinitrobenzene (CDNB) was examined for use in rapid detection of expression targeted to inclusion bodies and for the identification of inclusion body proteins which can be solubilized in low concentrations of chaotropic agents. Applicability of the assay was evaluated by tracking protein expression of two GST-fused allergens of biopharmaceutical value in E. coli, GST-Der p 2 and GST-Mus a 5, both targeted to inclusion bodies.
PB  - Elsevier Science Bv, Amsterdam
T2  - Journal of Biotechnology
T1  - Employment of colorimetric enzyme assay for monitoring expression and solubility of GST fusion proteins targeted to inclusion bodies
VL  - 168
IS  - 4
SP  - 506
EP  - 510
DO  - 10.1016/j.jbiotec.2013.09.019
ER  - 

@article{
author = "Macinkovic, Igor S. and Abughren, Mohamed and Mrkić, Ivan and Grozdanović, Milica M. and Prodanović, Radivoje and Gavrović-Jankulović, Marija",
year = "2013",
abstract = "High levels of recombinant protein expression can lead to the formation of insoluble inclusion bodies. These complex aggregates are commonly solubilized in strong denaturants, such as 6-8 M urea, although, if possible, solubilization under milder conditions could facilitate subsequent refolding and purification of bioactive proteins. Commercially available GST-tag assays are designed for quantitative measurement of GST activity under native conditions. GST fusion proteins accumulated in inclusion bodies are considered to be undetectable by such assays. In this work, solubilization of recombinantly produced proteins was performed in 4 M urea. The activity of rGST was assayed in 2 M urea and it was shown that rGST preserves 85% of its activity under such denaturing conditions. A colorimetric GST activity assay with 1-chloro-2, 4-dinitrobenzene (CDNB) was examined for use in rapid detection of expression targeted to inclusion bodies and for the identification of inclusion body proteins which can be solubilized in low concentrations of chaotropic agents. Applicability of the assay was evaluated by tracking protein expression of two GST-fused allergens of biopharmaceutical value in E. coli, GST-Der p 2 and GST-Mus a 5, both targeted to inclusion bodies.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Journal of Biotechnology",
title = "Employment of colorimetric enzyme assay for monitoring expression and solubility of GST fusion proteins targeted to inclusion bodies",
volume = "168",
number = "4",
pages = "506-510",
doi = "10.1016/j.jbiotec.2013.09.019"
}

Macinkovic, I. S., Abughren, M., Mrkić, I., Grozdanović, M. M., Prodanović, R.,& Gavrović-Jankulović, M.. (2013). Employment of colorimetric enzyme assay for monitoring expression and solubility of GST fusion proteins targeted to inclusion bodies. in Journal of Biotechnology
Elsevier Science Bv, Amsterdam., 168(4), 506-510.
https://doi.org/10.1016/j.jbiotec.2013.09.019

Macinkovic IS, Abughren M, Mrkić I, Grozdanović MM, Prodanović R, Gavrović-Jankulović M. Employment of colorimetric enzyme assay for monitoring expression and solubility of GST fusion proteins targeted to inclusion bodies. in Journal of Biotechnology. 2013;168(4):506-510.
doi:10.1016/j.jbiotec.2013.09.019 .

Macinkovic, Igor S., Abughren, Mohamed, Mrkić, Ivan, Grozdanović, Milica M., Prodanović, Radivoje, Gavrović-Jankulović, Marija, "Employment of colorimetric enzyme assay for monitoring expression and solubility of GST fusion proteins targeted to inclusion bodies" in Journal of Biotechnology, 168, no. 4 (2013):506-510,
https://doi.org/10.1016/j.jbiotec.2013.09.019 . .

TY  - JOUR
AU  - Grozdanović, Milica M.
AU  - Drakulić, Branko J.
AU  - Gavrović-Jankulović, Marija
PY  - 2013
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1398
AB  - Background: Actinidin, a protease from kiwifruit, belongs to the C1 family of cysteine proteases. Cysteine proteases were found to be involved in many disease states and are valid therapeutic targets. Actinidin has a wide pH activity range and wide substrate specificity, which makes it a good model system for studying enzyme-substrate interactions. Methods: The influence of inhibitor (E-64) binding on the conformation of actinidin was examined by 2D PAGE, circular dichroism (CD) spectroscopy, hydrophobic ligand binding assay, and molecular dynamics simulations. Results: Significant differences were observed in electrophoretic mobility of proteolytically active and E-64-inhibited actinidin. CD spectrometry and hydrophobic ligand binding assay revealed a difference in conformation between active and inhibited actinidin. Molecular dynamics simulations showed that a loop defined by amino-acid residues 88-104 had greater conformational mobility in the inhibited enzyme than in the active one. During MD simulations, the covalently bound inhibitor was found to change its conformation from extended to folded, with the guanidino moiety approaching the carboxylate. Conclusions: Conformational mobility of actinidin changes upon binding of the inhibitor, leading to a sequence of events that enables water and ions to protrude into a newly formed cavity of the inhibited enzyme. Drastic conformational mobility of E-64, a common inhibitor of cysteine proteases found in many crystal structures stored in PDB, was also observed. General significance: The analysis of structural changes which occur upon binding of an inhibitor to a cysteine protease provides a valuable starting point for the future design of therapeutic agents.
PB  - Elsevier Science Bv, Amsterdam
T2  - Biochimica et Biophysica Acta: General Subjects
T1  - Conformational mobility of active and E-64-inhibited actinidin
VL  - 1830
IS  - 10
SP  - 4790
EP  - 4799
DO  - 10.1016/j.bbagen.2013.06.015
ER  - 

@article{
author = "Grozdanović, Milica M. and Drakulić, Branko J. and Gavrović-Jankulović, Marija",
year = "2013",
abstract = "Background: Actinidin, a protease from kiwifruit, belongs to the C1 family of cysteine proteases. Cysteine proteases were found to be involved in many disease states and are valid therapeutic targets. Actinidin has a wide pH activity range and wide substrate specificity, which makes it a good model system for studying enzyme-substrate interactions. Methods: The influence of inhibitor (E-64) binding on the conformation of actinidin was examined by 2D PAGE, circular dichroism (CD) spectroscopy, hydrophobic ligand binding assay, and molecular dynamics simulations. Results: Significant differences were observed in electrophoretic mobility of proteolytically active and E-64-inhibited actinidin. CD spectrometry and hydrophobic ligand binding assay revealed a difference in conformation between active and inhibited actinidin. Molecular dynamics simulations showed that a loop defined by amino-acid residues 88-104 had greater conformational mobility in the inhibited enzyme than in the active one. During MD simulations, the covalently bound inhibitor was found to change its conformation from extended to folded, with the guanidino moiety approaching the carboxylate. Conclusions: Conformational mobility of actinidin changes upon binding of the inhibitor, leading to a sequence of events that enables water and ions to protrude into a newly formed cavity of the inhibited enzyme. Drastic conformational mobility of E-64, a common inhibitor of cysteine proteases found in many crystal structures stored in PDB, was also observed. General significance: The analysis of structural changes which occur upon binding of an inhibitor to a cysteine protease provides a valuable starting point for the future design of therapeutic agents.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Biochimica et Biophysica Acta: General Subjects",
title = "Conformational mobility of active and E-64-inhibited actinidin",
volume = "1830",
number = "10",
pages = "4790-4799",
doi = "10.1016/j.bbagen.2013.06.015"
}

Grozdanović, M. M., Drakulić, B. J.,& Gavrović-Jankulović, M.. (2013). Conformational mobility of active and E-64-inhibited actinidin. in Biochimica et Biophysica Acta: General Subjects
Elsevier Science Bv, Amsterdam., 1830(10), 4790-4799.
https://doi.org/10.1016/j.bbagen.2013.06.015

Grozdanović MM, Drakulić BJ, Gavrović-Jankulović M. Conformational mobility of active and E-64-inhibited actinidin. in Biochimica et Biophysica Acta: General Subjects. 2013;1830(10):4790-4799.
doi:10.1016/j.bbagen.2013.06.015 .

Grozdanović, Milica M., Drakulić, Branko J., Gavrović-Jankulović, Marija, "Conformational mobility of active and E-64-inhibited actinidin" in Biochimica et Biophysica Acta: General Subjects, 1830, no. 10 (2013):4790-4799,
https://doi.org/10.1016/j.bbagen.2013.06.015 . .