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

IF 2.8 Q3 Biochemistry, Genetics and Molecular Biology

Journal of Genetic Engineering and Biotechnology Pub Date : 2025-06-17 DOI:10.1016/j.jgeb.2025.100509

Mentalla Motaz Abdellatif , Azza El Amir , Said Shalaby , Michael Kirschfink , Rola Nadeem , Dina Nadeem Abd-Elshafy , Mahmoud Mohamed Bahgat

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引用次数: 0

Abstract

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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人唾液中参与SARS-CoV-2发病机制的蛋白酶分析：年龄和性别的影响

背景sars - cov -2通过血管紧张素转换酶2 （ACE2）进入人体细胞。计算机实验也证明了SARS-CoV-2在细胞进入过程中可能与二肽基肽酶4 （DPP-4）结合。此外，基质金属蛋白酶2 （MMP-2）参与了SARS-CoV-2细胞融合过程。目的研究不同年龄人群中SARS-CoV-2感染相关酶的表达和活性谱，探讨其对COVID-19结局的可能影响。方法采用酶谱法对不同年龄组的唾液进行MMP-2和明胶酶的酶谱分析。H-Arg-Pro-pNA底物裂解产生的酶活性使用微孔板阅读器进行测定。采用qRT-PCR和ELISA法对ACE2的转录和翻译进行定量分析。采用统计学分析来确定是否存在显著差异。结果酶谱分析显示，明胶酶在~ 180 ~ 30 kDa范围内具有不同的活性，在~ 96、72、66、60和40 kDa处显示出不同的条带。性别间MMP-2、DPP-4、DPP-4/ACE2活性、ACE2蛋白和mRNA水平的p值分别为0.0142、0.1056、0.1859、0.6411和0.4797，与年龄的相关系数r分别为0.1377、0.4185、0.0597、- 0.0993和- 0.0073。ACE2蛋白与具有DPP-4活性的ACE2 mRNA之间的r值分别为- 0.0668和0.1935。DPP-4活性和ACE2蛋白与MMP-2活性的差值分别为- 0.0506和0.0898。结果表明：(1)男性的MMP-2活性较高。(2) DPP-4活性与年龄呈正相关。(3) DPP-4活性、ACE2蛋白、mRNA水平的性别差异无统计学意义。(4) MMP-2活性、ACE2表达的年龄差异无统计学意义。(5)酶间不存在相关性。结论MMP-2和DPP-4的活性差异提示它们可能是男性和老年人对SARS-CoV-2感染易感性和COVID-19严重程度的危险因素。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Genetic Engineering and Biotechnology Biochemistry, Genetics and Molecular Biology-Biotechnology

CiteScore

5.70

自引率

5.70%

发文量

159

审稿时长

16 weeks

期刊介绍： Journal of genetic engineering and biotechnology is devoted to rapid publication of full-length research papers that leads to significant contribution in advancing knowledge in genetic engineering and biotechnology and provide novel perspectives in this research area. JGEB includes all major themes related to genetic engineering and recombinant DNA. The area of interest of JGEB includes but not restricted to: •Plant genetics •Animal genetics •Bacterial enzymes •Agricultural Biotechnology, •Biochemistry, •Biophysics, •Bioinformatics, •Environmental Biotechnology, •Industrial Biotechnology, •Microbial biotechnology, •Medical Biotechnology, •Bioenergy, Biosafety, •Biosecurity, •Bioethics, •GMOS, •Genomic, •Proteomic JGEB accepts