Profiling of proteases involved in SARS-CoV-2 pathogenesis in human saliva: Influence of age and gender

Background: SARS-CoV-2 enters human cells via angiotensin-converting enzyme 2 (ACE2). In silico experiments also demonstrated possible SARS-CoV-2 binding to dipeptidyl peptidase 4 (DPP-4) during cell entry. Moreover, matrix metalloproteinase 2 (MMP-2) is involved in the SARS-CoV-2 cell fusion proces...

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Main Authors: Mentalla Motaz Abdellatif, Azza El Amir, Said Shalaby, Michael Kirschfink, Rola Nadeem, Dina Nadeem Abd-Elshafy, Mahmoud Mohamed Bahgat
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:Journal of Genetic Engineering and Biotechnology
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Online Access:http://www.sciencedirect.com/science/article/pii/S1687157X25000538
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author Mentalla Motaz Abdellatif
Azza El Amir
Said Shalaby
Michael Kirschfink
Rola Nadeem
Dina Nadeem Abd-Elshafy
Mahmoud Mohamed Bahgat
author_facet Mentalla Motaz Abdellatif
Azza El Amir
Said Shalaby
Michael Kirschfink
Rola Nadeem
Dina Nadeem Abd-Elshafy
Mahmoud Mohamed Bahgat
author_sort Mentalla Motaz Abdellatif
collection DOAJ
description Background: SARS-CoV-2 enters human cells via angiotensin-converting enzyme 2 (ACE2). In silico experiments also demonstrated possible SARS-CoV-2 binding to dipeptidyl peptidase 4 (DPP-4) during cell entry. Moreover, matrix metalloproteinase 2 (MMP-2) is involved in the SARS-CoV-2 cell fusion process. Aims: To study expression and activity profiles of implicated enzymes in SARS-CoV-2 infection in various age groups of both genders and discuss the possible impact of this on COVID-19 outcome. Methods: Saliva from different age groups was used to profile MMP-2 and gelatinase enzymes by zymography. Enzyme activities resulting from the cleavage of the H-Arg-Pro-pNA substrate were measured using a microplate reader. ACE2 transcription and translation were quantified using qRT-PCR and ELISA. Statistical analysis was used to identify the presence of significant differences. Results: Zymography demonstrated different gelatinase activities in a range of ∼180–30 kDa and showed distinct bands at ∼96, 72, 66, 60, and 40 kDa. Comparing MMP-2, DPP-4, DPP-4/ACE2 activities, ACE2 protein, and mRNA levels between genders showed p-values of 0.0142, 0.1056, 0.1859, 0.6411, and 0.4797, whereas correlations with age showed correlation coefficient (r) values of 0.1377, 0.4185, 0.0597, −0.0993, and −0.0073, respectively. The r values between ACE2 protein and ACE2 mRNA with DPP-4 activity were −0.0668 and 0.1935, respectively. The values between both DPP-4 activity and ACE2 protein with MMP-2 activity were −0.0506 and 0.0898, respectively. Results reflected (1) MMP-2 activity is higher among males. (2) DPP-4 activity positively correlates with age. (3) Gender differences in DPP-4 activity, ACE2 protein, and mRNA levels were not statistically significant. (4) Age differences in MMP-2 activity, ACE2 expression were not significant. (5) Inter-enzyme correlations were not observed. Conclusions: The differential activity of both MMP-2 and DPP-4 highlights their potential as risk factors for susceptibility to SARS-CoV-2 infection and COVID-19 severity in males and older groups.
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spelling doaj-art-91eee30aeca74f5e933a3df1e6bc21472025-08-24T05:11:42ZengElsevierJournal of Genetic Engineering and Biotechnology1687-157X2025-09-0123310050910.1016/j.jgeb.2025.100509Profiling of proteases involved in SARS-CoV-2 pathogenesis in human saliva: Influence of age and genderMentalla Motaz Abdellatif0Azza El Amir1Said Shalaby2Michael Kirschfink3Rola Nadeem4Dina Nadeem Abd-Elshafy5Mahmoud Mohamed Bahgat6Department of Therapeutic Chemistry, Institute of Pharmaceutical and Drug Industries Research, the National Research Centre, Dokki, Giza 12311, Egypt; Research Group Immune- and Biomarkers for Infection, the Centre of Excellence for Advanced Sciences, the National Research Centre, Giza 12311, EgyptZoology Department, Faculty of Science, Cairo University, Giza 12613, EgyptDepartment of Complementary and Alternative Medicine, Medical Research Division, National Research Centre, Dokki, 12622 Giza, EgyptInstitute of Immunology, Ruprecht Karls University of Heidelberg, Heidelberg, GermanyDepartment of Therapeutic Chemistry, Institute of Pharmaceutical and Drug Industries Research, the National Research Centre, Dokki, Giza 12311, Egypt; Research Group Immune- and Biomarkers for Infection, the Centre of Excellence for Advanced Sciences, the National Research Centre, Giza 12311, EgyptResearch Group Immune- and Biomarkers for Infection, the Centre of Excellence for Advanced Sciences, the National Research Centre, Giza 12311, Egypt; Environmental Virology Laboratory, Department of Water Pollution Research, Institute of Environmental Research and Climate Changes, the National Research Centre, Dokki, Giza 12311, EgyptDepartment of Therapeutic Chemistry, Institute of Pharmaceutical and Drug Industries Research, the National Research Centre, Dokki, Giza 12311, Egypt; Research Group Immune- and Biomarkers for Infection, the Centre of Excellence for Advanced Sciences, the National Research Centre, Giza 12311, Egypt; Egypt Center for Research and Regenerative Medicine, Al Abbassia, Cairo 11517, Egypt; Corresponding author at: Department of Therapeutic Chemistry, Institute of Pharmaceutical and Drug Industries Research, the National Research Centre, Dokki, Giza 12311, Egypt.Background: SARS-CoV-2 enters human cells via angiotensin-converting enzyme 2 (ACE2). In silico experiments also demonstrated possible SARS-CoV-2 binding to dipeptidyl peptidase 4 (DPP-4) during cell entry. Moreover, matrix metalloproteinase 2 (MMP-2) is involved in the SARS-CoV-2 cell fusion process. Aims: To study expression and activity profiles of implicated enzymes in SARS-CoV-2 infection in various age groups of both genders and discuss the possible impact of this on COVID-19 outcome. Methods: Saliva from different age groups was used to profile MMP-2 and gelatinase enzymes by zymography. Enzyme activities resulting from the cleavage of the H-Arg-Pro-pNA substrate were measured using a microplate reader. ACE2 transcription and translation were quantified using qRT-PCR and ELISA. Statistical analysis was used to identify the presence of significant differences. Results: Zymography demonstrated different gelatinase activities in a range of ∼180–30 kDa and showed distinct bands at ∼96, 72, 66, 60, and 40 kDa. Comparing MMP-2, DPP-4, DPP-4/ACE2 activities, ACE2 protein, and mRNA levels between genders showed p-values of 0.0142, 0.1056, 0.1859, 0.6411, and 0.4797, whereas correlations with age showed correlation coefficient (r) values of 0.1377, 0.4185, 0.0597, −0.0993, and −0.0073, respectively. The r values between ACE2 protein and ACE2 mRNA with DPP-4 activity were −0.0668 and 0.1935, respectively. The values between both DPP-4 activity and ACE2 protein with MMP-2 activity were −0.0506 and 0.0898, respectively. Results reflected (1) MMP-2 activity is higher among males. (2) DPP-4 activity positively correlates with age. (3) Gender differences in DPP-4 activity, ACE2 protein, and mRNA levels were not statistically significant. (4) Age differences in MMP-2 activity, ACE2 expression were not significant. (5) Inter-enzyme correlations were not observed. Conclusions: The differential activity of both MMP-2 and DPP-4 highlights their potential as risk factors for susceptibility to SARS-CoV-2 infection and COVID-19 severity in males and older groups.http://www.sciencedirect.com/science/article/pii/S1687157X25000538Angiotensin converting enzyme 2Dipeptidyl peptidase 4Matrix metalloproteases 2SARS-CoV-2COVID-19 severityActivities
spellingShingle Mentalla Motaz Abdellatif
Azza El Amir
Said Shalaby
Michael Kirschfink
Rola Nadeem
Dina Nadeem Abd-Elshafy
Mahmoud Mohamed Bahgat
Profiling of proteases involved in SARS-CoV-2 pathogenesis in human saliva: Influence of age and gender
Journal of Genetic Engineering and Biotechnology
Angiotensin converting enzyme 2
Dipeptidyl peptidase 4
Matrix metalloproteases 2
SARS-CoV-2
COVID-19 severity
Activities
title Profiling of proteases involved in SARS-CoV-2 pathogenesis in human saliva: Influence of age and gender
title_full Profiling of proteases involved in SARS-CoV-2 pathogenesis in human saliva: Influence of age and gender
title_fullStr Profiling of proteases involved in SARS-CoV-2 pathogenesis in human saliva: Influence of age and gender
title_full_unstemmed Profiling of proteases involved in SARS-CoV-2 pathogenesis in human saliva: Influence of age and gender
title_short Profiling of proteases involved in SARS-CoV-2 pathogenesis in human saliva: Influence of age and gender
title_sort profiling of proteases involved in sars cov 2 pathogenesis in human saliva influence of age and gender
topic Angiotensin converting enzyme 2
Dipeptidyl peptidase 4
Matrix metalloproteases 2
SARS-CoV-2
COVID-19 severity
Activities
url http://www.sciencedirect.com/science/article/pii/S1687157X25000538
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